Re: [Histonet] Help to interview new employees for the first time

[Sheeder, Christopher via Histonet]

Hi Blanca,
Hiring has become more difficult in recent years. Most employers now only 
verify past employment. They cannot divulge any corrective actions, performance 
issues or firings.
References are hand-picked by the applicant so you don't get the whole picture 
there either.
I typically ask them about their experiences in histology. Obstacles they have 
overcome, how to handle difficult customers (angry physicians) etc.
If you can give the other techs a chance to ask the candidate questions, great, 
otherwise see what questions your techs want to know.
I fully agree with Terri Braud's response. Do not have them perform any type of 
function. That is the purpose of the probationary period.
The best interview question that was ever asked of me..."Name 12 uses for a 
pencil".
Seems silly, but it's not about the answer, it's about demonstrating your 
creative problem solving skills. (it was tough, but I came up with 12!)
Best of luck!

Christopher Sheeder, HT(ASCP)QIHC
Pathology Manager | Department of Laboratories
Seattle Children's Hospital

-Original Message-
From: Blanca Lopez 
Sent: Thursday, September 12, 2019 11:48 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Help to interview new employees for the first time

I am going to interview people for Histotech position for the first time...what 
are the best questions to ask? How do I prepare myself? what is the I need to 
know that they are the best one? What should I ask or choose? Is good to put 
them in action like cutting or staining to check on their skills or what are 
your recommendations? thank you for your help

Blanca Lopez HT (ASCP)cm
Senior Histotechnologist
UT Southwestern Medical Center
Harold C. Simmons Comprehensive Cancer Center UTSTR Biorepository Tissue Lab
6000 Harry Hines Blvd NB5.102
Dallas, Texas 75390
214-648-7598
blanca.lo...@utsouthwestern.edu





UT Southwestern


Medical Center



The future of medicine, today.


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Re: [Histonet] Help to interview new employees for the first time

[Patrick Laurie via Histonet]

I'm also a fan of having any fellow employees available ask questions.  I
have my staff limit it to histology related subjects, but my thought is if
they are going to be the ones working directly with them, it helps to have
their opinions.

Patrick Laurie(HT)ASCP QIHC

Histology Manager

Celligent Diagnostics, LLC

101 East W.T. Harris Blvd  | Suite 1212 | Charlotte, NC 28262

Work: 704-970-3300  Cell: 704-266-0869


On Fri, Sep 13, 2019 at 10:40 AM Anne Murvosh via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> I'm not sure if this is an option, but we used to melt down old blocks and
> make the HT embed and cut them. This showed us how good and quick they were
> and if they actually new what they were doing. Anne
>
> -Original Message-
> From: Blanca Lopez via Histonet 
> Sent: Thursday, September 12, 2019 11:48 AM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Help to interview new employees for the first time
>
> I am going to interview people for Histotech position for the first
> time...what are the best questions to ask? How do I prepare myself? what is
> the I need to know that they are the best one? What should I ask or choose?
> Is good to put them in action like cutting or staining to check on their
> skills or what are your recommendations? thank you for your help
>
> Blanca Lopez HT (ASCP)cm
> Senior Histotechnologist
> UT Southwestern Medical Center
> Harold C. Simmons Comprehensive Cancer Center UTSTR Biorepository Tissue
> Lab
> 6000 Harry Hines Blvd NB5.102
> Dallas, Texas 75390
> 214-648-7598
> blanca.lo...@utsouthwestern.edu
>
>
>
> 
>
> UT Southwestern
>
>
> Medical Center
>
>
>
> The future of medicine, today.
>
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Re: [Histonet] Help to interview new employees for the first time

[Anne Murvosh via Histonet]

I'm not sure if this is an option, but we used to melt down old blocks and make 
the HT embed and cut them. This showed us how good and quick they were and if 
they actually new what they were doing. Anne

-Original Message-
From: Blanca Lopez via Histonet  
Sent: Thursday, September 12, 2019 11:48 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Help to interview new employees for the first time

I am going to interview people for Histotech position for the first time...what 
are the best questions to ask? How do I prepare myself? what is the I need to 
know that they are the best one? What should I ask or choose? Is good to put 
them in action like cutting or staining to check on their skills or what are 
your recommendations? thank you for your help

Blanca Lopez HT (ASCP)cm
Senior Histotechnologist
UT Southwestern Medical Center
Harold C. Simmons Comprehensive Cancer Center UTSTR Biorepository Tissue Lab
6000 Harry Hines Blvd NB5.102
Dallas, Texas 75390
214-648-7598
blanca.lo...@utsouthwestern.edu





UT Southwestern


Medical Center



The future of medicine, today.

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Re: [Histonet] Help to interview new employees for the first time

[Erin McCarthy via Histonet]

Hi Blanca,

I think practical skills tests are a great idea. BUT I also think you need
to make sure that having someone that isn't an employee doing tasks in the
lab is a responsibility your lab would willingly take on if something were
to happen. If the candidate cut themselves and was litigious they could go
after your lab. I have not heard of it happening, but I know in this day
and age it can be a risk. Otherwise I try to ask probing questions -
usually things that cannot be answered with Yes or No. Also, I try to ask
about things you really want to know - how do they manage stress, do they
have troubleshooting experience, if so can they explain the problem and how
they solved it. Are they intuitive enough, that looking back on a
troubleshooting or stressful event can they identify what they would have
done differently? I want to make sure that the people I hire can manage
stress, and think through an issue. I also try to ask them what they feel
makes a good tech, and how they think their coworkers would describe them.
It gets them thinking about the team, not just how they see themselves.

I hope this helps!

On Thu, Sep 12, 2019 at 2:06 PM Blanca Lopez via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> I am going to interview people for Histotech position for the first
> time...what are the best questions to ask? How do I prepare myself? what is
> the I need to know that they are the best one? What should I ask or choose?
> Is good to put them in action like cutting or staining to check on their
> skills or what are your recommendations? thank you for your help
>
> Blanca Lopez HT (ASCP)cm
> Senior Histotechnologist
> UT Southwestern Medical Center
> Harold C. Simmons Comprehensive Cancer Center
> UTSTR Biorepository Tissue Lab
> 6000 Harry Hines Blvd NB5.102
> Dallas, Texas 75390
> 214-648-7598
> blanca.lo...@utsouthwestern.edu
>
>
>
> 
>
> UT Southwestern
>
>
> Medical Center
>
>
>
> The future of medicine, today.
>
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>


-- 

Erin McCarthy, HT (ASCP)
Histology Supervisor

Tempus Labs
600 W. Chicago Ave.
Chicago IL 60654
Cell: (708)269-8610

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Re: [Histonet] HELP Path requesting +PAP stain control

[Terri Braud via Histonet]

On Date: Mon, 23 Jul 2018 18:31:48 -0400
From: Mary Ann 
Subject: [Histonet] Positive PAP
Mary Ann wrote "Help! My pathologist has asked that a positive patient be run 
down with our PAP stain for QC.
Point me to a reference to counter this request."

Hi Mary Ann - First of all, my sympathies.  This is the kind of craziness that 
can give a pathologist a bad name.  Secondly, what does he call a patient 
positive?  Positive for what?  LOL, JK.  In response to your question, here are 
the ONLY 2 requirements for Cytology stain QC, straight from the latest CAP 
list.  See below.  As one can see, nowhere does it require any type of patient 
control, only a documented assessment of the stain quality, on "actual case 
material"  CAPs words, not mine. Good Luck! Terri
__

**REVISED**   08/21/2017
CYP.03925   Stain AssessmentPhase I
Cytology stains are assessed at least annually to ensure their proper 
storage and acceptable quality.
NOTE: Cytology stains undergoing a daily technical quality review are exempt 
from an annual assessment.
Most stains used in the cytology laboratory are not subject to outdating, so 
that assignment of expiration dates may have no meaning.  The acceptable 
performance of such stains must be confirmed at least annually by technical 
assessment on actual case material, and as part of the evaluation of 
cytopathology cases. Where applicable, expiration dates assigned by a 
manufacturer must be observed.
Evidence of Compliance:
✓   Written procedure for stain assessment AND
✓   Records of assessment of appropriate quality of each cytology stain in 
use

CYP.04300   Daily QCPhase II
Daily QCPhase II
There are records of daily review of the technical quality of cytologic 
preparations by the pathologist or supervisory-level cytotechnologist.
NOTE:  The technical quality of cytologic preparations must be checked daily 
(on days processing occurs). This includes checking all stains for predicted 
staining characteristics each day of use. This check must include all of the 
types of preparations seen that day such as cytospins, cell blocks, and liquid 
based preparations.
If preparation and staining is performed by a different laboratory, there must 
be a procedure for the laboratory performing the preparation and staining to 
verify the acceptability of the quality of preparations and the acceptability 
of controls (if needed) before transfer.  Records of this verification must be 
readily available to the laboratory performing interpretations.  There should 
also be a mechanism for feedback from the interpreting laboratory to the 
laboratory that prepared the slides of any issues with the preparations.
_

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874
Care, Comfort, and Heal

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Re: [Histonet] Help with preparing slides of baby guppies

[J B via Histonet]

Jenn,

Trying to fully understand. You need someone to prepare these slides again
for you?  Great quality work, you provide the specimen?  Let me know, I
would love to learn more.

Sincerely,

JB

On Wed, May 31, 2017, 9:13 AM Dearolf, Jenn via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Hello!
>
> My name is Jenn Dearolf, and I am a professor in the Biology Department at
> Hendrix College, a small liberal arts college in Conway, AR.  I have
> written this list before to get advice about how to prevent freezing
> artifact in small muscle samples (Thanks!), but today, I have a completely
> different need.
>
> In our Zoology course at Hendrix, we use slides that have numerous serial
> sections (7 to 10 microns thickness) of a baby guppy on them that have been
> stained with H & E.  And, a box of slides, ranging from 8 to 12 slides, is
> the entire guppy sectioned from the tip of its nose to the tip of its
> tail.  However, these slides are very old, and over the years, the mounting
> media has pulled away from the sections.  In addition, numerous slides have
> been broken or lost.
>
> Unfortunately, no one in my Department now has the necessary skills to
> produce these slides for our students.  I was wondering if anyone on the
> list knew of a facility that could produce these slides.  I think we should
> be able to provide the baby guppies.  I would just have to get approval
> from our IACUC.
>
> I also have a very old paper that discusses the steps that were necessary
> to produce the slides.  I am happy to share the methodology with anyone
> that could help us, and we could discuss what we would need to do at
> Hendrix and which steps would need to be performed at your facility.
>
> I appreciate any advice folks are willing to share.  Thanks for your time
> and consideration.
>
> Sincerely,
> Jenn
>
>
> Jennifer Dearolf, Ph.D.
> Professor and Chair
> Biology Department
> Hendrix College
> 1600 Washington Ave.
> Conway, AR 72032
> (501) 450-4530 (office)
>
>
>
>
>
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Re: [Histonet] help!!

[Tony Henwood (SCHN) via Histonet]

You can use DAB but the issue of endogenous peroxidase will rear its ugly head.

And I suppose IF being around since 1955, when Mellors first applied the 
technique to renal tissue, it has a long history of diagnostic application that 
is hard to replace.

There are other enzymes that could be used that, not being present in human 
tissue, would not require endogenous enzyme blocking for example glucose 
oxidase. The advantage of this enzyme is that there is no endogenous glucose 
oxidase activity in mammalian tissues.

Weening, J. J., & Jennette, J. C. (2012). Historical milestones in renal 
pathology. Virchows Archiv, 461(1), 3-11.

Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
Principal Scientist, the Children’s Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney
Tel: 612 9845 3306
Fax: 612 9845 3318
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

From: Allan Wang [mailto:alla...@gmail.com]
Sent: Tuesday, 18 April 2017 5:51 PM
To: Tony Henwood (SCHN)
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] help!!

Tim and Tony,

Why couldn't DAB be used on frozen sections in your example?

Allan

On Mon, Apr 17, 2017 at 9:03 PM, Tony Henwood (SCHN) via Histonet 
<histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>> 
wrote:
Hi Bianca,
Well for most Pathology departments, Immunofluorescence (IF) is used for Renal 
and Skin biopsies; looking for human Immunoglobulin (Igs) deposition on 
basement membranes. The advantage here is not so much the fluorescence, but 
that we use unfixed frozen sections. The buffer rinse before antibody 
application, removes un-bound serum immunoglobulins, leaving any pathological 
bound Igs for the IF antibody to bind to. This gives a clean result.

If one would do IF on formalin-fixed paraffin sections of renal or skin 
biopsies, you would find heavy background due to the fixative cross-linking 
serum Igs to tissue and cells (which would usually be removed by the buffer 
rinse if unfixed frozen sections were used - see above).

IF, apart from being a historic method, also does not suffer from endogenous 
peroxidase that would need to be blocked if peroxidase was used in place of 
fluorescence.



Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
Principal Scientist, the Children's Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney
Tel: 612 9845 3306
Fax: 612 9845 3318
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

-Original Message-
From: Blanca Lopez via Histonet 
[mailto:histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>]
Sent: Thursday, 13 April 2017 11:10 PM
To: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>
Subject: [Histonet] help!!
Hello!
I just need a help with a simple question...Is anyone can explain me what is 
the purpose between performing immunohistochemistry and Immunofluorescence?
Thanks  :)

Blanca Lopez
Histotech (ASCP)
UTSW Tissue Resource K1.210
Simmons Comprehensive Cancer Center
UT Southwestern Medical Center
Telephone: 214-648-7598
Email: blanca.lo...@utsouthwestern.edu<mailto:blanca.lo...@utsouthwestern.edu>




UT Southwestern


Medical Center



The future of medicine, today.

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Re: [Histonet] help!!

[Allan Wang via Histonet]

Tim and Tony,

Why couldn't DAB be used on frozen sections in your example?

Allan

On Mon, Apr 17, 2017 at 9:03 PM, Tony Henwood (SCHN) via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Hi Bianca,
> Well for most Pathology departments, Immunofluorescence (IF) is used for
> Renal and Skin biopsies; looking for human Immunoglobulin (Igs) deposition
> on basement membranes. The advantage here is not so much the fluorescence,
> but that we use unfixed frozen sections. The buffer rinse before antibody
> application, removes un-bound serum immunoglobulins, leaving any
> pathological bound Igs for the IF antibody to bind to. This gives a clean
> result.
>
> If one would do IF on formalin-fixed paraffin sections of renal or skin
> biopsies, you would find heavy background due to the fixative cross-linking
> serum Igs to tissue and cells (which would usually be removed by the buffer
> rinse if unfixed frozen sections were used - see above).
>
> IF, apart from being a historic method, also does not suffer from
> endogenous peroxidase that would need to be blocked if peroxidase was used
> in place of fluorescence.
>
>
>
> Regards
> Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
> Principal Scientist, the Children's Hospital at Westmead
> Adjunct Fellow, School of Medicine, University of Western Sydney
> Tel: 612 9845 3306
> Fax: 612 9845 3318
> Pathology Department
> the children's hospital at westmead
> Cnr Hawkesbury Road and Hainsworth Street, Westmead
> Locked Bag 4001, Westmead NSW 2145, AUSTRALIA
>
> -Original Message-
> From: Blanca Lopez via Histonet [mailto:histonet@lists.utsouthwestern.edu]
> Sent: Thursday, 13 April 2017 11:10 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] help!!
>
> Hello!
> I just need a help with a simple question...Is anyone can explain me what
> is the purpose between performing immunohistochemistry and
> Immunofluorescence?
> Thanks  :)
>
> Blanca Lopez
> Histotech (ASCP)
> UTSW Tissue Resource K1.210
> Simmons Comprehensive Cancer Center
> UT Southwestern Medical Center
> Telephone: 214-648-7598
> Email: blanca.lo...@utsouthwestern.edu
>
>
> 
>
> UT Southwestern
>
>
> Medical Center
>
>
>
> The future of medicine, today.
>
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> This message is intended for the addressee named and may contain
> confidential information. If you are not the intended recipient, please
> delete it and notify the sender.
>
> Views expressed in this message are those of the individual sender, and
> are not necessarily the views of NSW Health or any of its entities.
>
>
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Re: [Histonet] help!!

[Tony Henwood (SCHN) via Histonet]

Hi Bianca,
Well for most Pathology departments, Immunofluorescence (IF) is used for Renal 
and Skin biopsies; looking for human Immunoglobulin (Igs) deposition on 
basement membranes. The advantage here is not so much the fluorescence, but 
that we use unfixed frozen sections. The buffer rinse before antibody 
application, removes un-bound serum immunoglobulins, leaving any pathological 
bound Igs for the IF antibody to bind to. This gives a clean result.

If one would do IF on formalin-fixed paraffin sections of renal or skin 
biopsies, you would find heavy background due to the fixative cross-linking 
serum Igs to tissue and cells (which would usually be removed by the buffer 
rinse if unfixed frozen sections were used - see above).

IF, apart from being a historic method, also does not suffer from endogenous 
peroxidase that would need to be blocked if peroxidase was used in place of 
fluorescence.



Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Principal Scientist, the Children's Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-Original Message-
From: Blanca Lopez via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Thursday, 13 April 2017 11:10 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] help!!

Hello!
I just need a help with a simple question...Is anyone can explain me what is 
the purpose between performing immunohistochemistry and Immunofluorescence?
Thanks  :)

Blanca Lopez
Histotech (ASCP)
UTSW Tissue Resource K1.210
Simmons Comprehensive Cancer Center
UT Southwestern Medical Center
Telephone: 214-648-7598
Email: blanca.lo...@utsouthwestern.edu




UT Southwestern


Medical Center



The future of medicine, today.

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Re: [Histonet] help!!

[Caroline Miller via Histonet]

Blanca,
Here are my feelings on this, and I am sure a lot of other folks have feels
here too, so please chime in.

1 - I feel that most clinical labs are more on the IHC bandwagon and
research labs are IF (with the exception of IgG staining in kidney biopsies
or bullous disease in skin- which is because the antibodies don't like
formalin fixing (if this is now wrong I am sorry, I haven't been in a
clinical lab in quite a while). Research labs are often also working with
genetically encoded fluorophores such as GFP, YFP, mCherry
2 - Formalin fixation (especially over fixation) can often lead to a large
amount of autofluorescence in the 488 region, which is a common place for
secondary antibodies and also GFP. Research labs have a lot more control
over their fixation protocols.
3 - The microscopes commonly available to clinical labs are bright field
scopes and in research labs fluorescent scopes
4 - Fluorescence can provide more contrast to a positively localized
fluorophore, but sometimes at the detriment of viewing the overall
morphology of the tissue like you get with bright field IHC and a nuclear
counterstain.
5 - Research lab protocols are often very 'experimental' and can lead to
increased tissue damage, which again is not viewed under the fluorescence
microscope (as much). Clinical labs have lots of experience and also
defined protocols that work well in the IHC / bright field space.
6 - the only real difference is the detection method, you can use any
primary antibody with either ABC/ impress / enzyme based methods or with
fluorophore conjugated secondaries.

So, in short - no *real* reason, but mainly that is the way things shook
out.

I could go on about researchers not understanding how to take photos on a
bright field scopes too, but that is too broad a statement, but as a core
director I saw them being more comfortable with the fluorescent methods :)

mills




On Thu, Apr 13, 2017 at 6:09 AM, Blanca Lopez via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Hello!
> I just need a help with a simple question...Is anyone can explain me what
> is the purpose between performing immunohistochemistry and
> Immunofluorescence?
> Thanks  :)
>
> Blanca Lopez
> Histotech (ASCP)
> UTSW Tissue Resource K1.210
> Simmons Comprehensive Cancer Center
> UT Southwestern Medical Center
> Telephone: 214-648-7598
> Email: blanca.lo...@utsouthwestern.edu
>
>
> 
>
> UT Southwestern
>
>
> Medical Center
>
>
>
> The future of medicine, today.
>
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>



-- 
Caroline Miller (mills)
Director of Histology
3Scan.com
415 2187297
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Re: [Histonet] help!!

[Morken, Timothy via Histonet]

Blanca, immunofluorescence (IF) is a subset of immunochemistry. 
Immunohistochemistry is also a subset of immunochemistry. There is some overlap 
between the two.

Immunohistochemistry denotes  immunochemistry done on tissue sections 
("-histo-" =" tissue"). But we can also use other enzymes to label the 
antibodies for immunohistochemistry (peroxidase, alkaline phosphatase, etc).


IF is just one of many methods of labeling the antibodies with a visual label. 
Others are peroxidase and alkaline phosphatase.


Generally IF is done on "fresh" cells or tissue. For tissue it is normally 
frozen tissue. 

IF can be done on cells (ie, immunocytochemistry) either on slides (smears, 
various preparations) or in solution as with flow cytometry - the cells are 
labeled with fluorescent-labeled antibodies and sorted by color (or no color). 

Generally the IF method is faster to perform because there is no processing 
beyond freezing the tissue. In the past IF was also more sensitive due to dark 
field microscopy in the fluorescence microscope. With the advent of various 
methods to amplify the signal (avidin -biotin, polymers with multiple enzymes) 
the peroxidase methods are just as sensitive, if not more so.

But fresh or frozen tissue has the advantage of the epitopes remaining unfixed, 
especially by formalin - which can mask the antigen from the antibody. Some 
antibodies do not work well on formalin-fixed tissue, even if antigen retrieval 
is used, so frozen tissue or cells are the best option. 



Tim Morken
Pathology Site Manager, Parnassus 
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center





-Original Message-
From: Blanca Lopez via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Thursday, April 13, 2017 6:10 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] help!!

Hello!
I just need a help with a simple question...Is anyone can explain me what is 
the purpose between performing immunohistochemistry and Immunofluorescence?
Thanks  :)

Blanca Lopez
Histotech (ASCP)
UTSW Tissue Resource K1.210
Simmons Comprehensive Cancer Center
UT Southwestern Medical Center
Telephone: 214-648-7598
Email: blanca.lo...@utsouthwestern.edu




UT Southwestern


Medical Center



The future of medicine, today.

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Re: [Histonet] Help with causes or theory behind failure of nuclear counterstaining after IHC!

[WILLIAM DESALVO via Histonet]

Maria, I. Think you may have a pH issue. High pH results in reduction of 
protons, H+, effects dye structure and can cause light to no staining after 
bluing. If you are using hematoxylin w/ aluminum, most popular, decreased pH = 
decreased intensity. Acid breaks the Al+3. Check your ph throughout the 
process. Sounds like something has changed. Good luck.

Sent from my Windows Phone

From: Morken, Timothy via Histonet<mailto:histonet@lists.utsouthwestern.edu>
Sent: ‎1/‎4/‎2016 9:20 AM
To: Maria Mejia<mailto:mbmph...@gmail.com>
Cc: Histonet<mailto:histonet@lists.utsouthwestern.edu>
Subject: Re: [Histonet] Help with causes or theory behind failure of nuclear 
counterstaining after IHC!

Maria,

If the counterstain is good when done before IHC stain and poor after it sounds 
like proteins are being extracted during the IHC processing and staining. Have 
you tried staining sections after each step of the IHC process to isolate the 
point the stain becomes weak?

Tim Morken
Pathology Site Manager, Parnassus
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center



-Original Message-
From: Maria Mejia via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Sunday, January 03, 2016 8:29 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Help with causes or theory behind failure of nuclear 
counterstaining after IHC!

Happy New Year Everyone,

I'm the lead histologist working in an IHC based research lab focused on early 
stages of Alzheimer's disease.  We work on paraffin sections processed & cut 
from 600um celloidin sections.  Including a lot of 60um cellodin sections from 
whole human brainstem.

For years, everything has going good regarding counterstaining after single & 
double IHC staining on 60um free-floating sections.  However for the past two 
months we've struggled to achieve good visible counterstaining on IHC sections 
to count the stained neurons - to see clearly the nucleus & nucleolus!

For a number of years, Gallocyanine was our choice of counterstain after IHC.  
Now, it's NOT working (neurons not stained visible enough to count).  We've 
also tried cresyl violet
counterstain -  staining too weak!   In both counterstains, we modified
the staining protocols quite a number of times to get good visible staining - 
nothing!!!

Strangle because we get lovely counterstained neurons with NO IHC staining on 
our 60um sections, but as soon as we take the sections through the IHC protocol 
e.g. antigen retrieval, antibodies & chromogens - counterstain is too weak!  
Our paraffin IHC sections work & look wonderful!

Now, my PI wants to try methyl green counterstain, however I think we'll have 
the same problem.
Here's what I need help with:

1) Can someone please explain the reason or theory  behind the failure of 
counterstain uptake by cells such as human neurons on 60um celloidin sections?

2) Can anyone please offer staining protocols that use alternative dehydration 
& clearing reagents.
I've been using alcohols dehydration (96% & 100%) without success as well as 
clearing with xylene - which hardens the tissue if left too long in this 
reagent.

 I was thinking of perhaps using acetone instead of alcohols & maybe using a 
methyl salicylate or chloroform.  Thoughts anyone?

I wish Dr Chris van der Loos was still with us.  I'd dearly like to hear from 
anyone who can help with this issue - Gayle Callis, Terry Johnson, Dr Hohn 
Kiernan et al.

Any assistance anyone can provide will be greatly appreciated!

Best
Maria Mejia
UCSF
Memory & Aging Department
San Francisco, CA
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Re: [Histonet] Help with causes or theory behind failure of nuclear counterstaining after IHC!

[Morken, Timothy via Histonet]

Maria, 

If the counterstain is good when done before IHC stain and poor after it sounds 
like proteins are being extracted during the IHC processing and staining. Have 
you tried staining sections after each step of the IHC process to isolate the 
point the stain becomes weak? 

Tim Morken
Pathology Site Manager, Parnassus 
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center



-Original Message-
From: Maria Mejia via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Sunday, January 03, 2016 8:29 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Help with causes or theory behind failure of nuclear 
counterstaining after IHC!

Happy New Year Everyone,

I'm the lead histologist working in an IHC based research lab focused on early 
stages of Alzheimer's disease.  We work on paraffin sections processed & cut 
from 600um celloidin sections.  Including a lot of 60um cellodin sections from 
whole human brainstem.  

For years, everything has going good regarding counterstaining after single & 
double IHC staining on 60um free-floating sections.  However for the past two 
months we've struggled to achieve good visible counterstaining on IHC sections 
to count the stained neurons - to see clearly the nucleus & nucleolus!  

For a number of years, Gallocyanine was our choice of counterstain after IHC.  
Now, it's NOT working (neurons not stained visible enough to count).  We've 
also tried cresyl violet
counterstain -  staining too weak!   In both counterstains, we modified
the staining protocols quite a number of times to get good visible staining - 
nothing!!!

Strangle because we get lovely counterstained neurons with NO IHC staining on 
our 60um sections, but as soon as we take the sections through the IHC protocol 
e.g. antigen retrieval, antibodies & chromogens - counterstain is too weak!  
Our paraffin IHC sections work & look wonderful!

Now, my PI wants to try methyl green counterstain, however I think we'll have 
the same problem.
Here's what I need help with:

1) Can someone please explain the reason or theory  behind the failure of 
counterstain uptake by cells such as human neurons on 60um celloidin sections?

2) Can anyone please offer staining protocols that use alternative dehydration 
& clearing reagents.
I've been using alcohols dehydration (96% & 100%) without success as well as 
clearing with xylene - which hardens the tissue if left too long in this 
reagent. 

 I was thinking of perhaps using acetone instead of alcohols & maybe using a 
methyl salicylate or chloroform.  Thoughts anyone?

I wish Dr Chris van der Loos was still with us.  I'd dearly like to hear from 
anyone who can help with this issue - Gayle Callis, Terry Johnson, Dr Hohn 
Kiernan et al.

Any assistance anyone can provide will be greatly appreciated!

Best
Maria Mejia
UCSF
Memory & Aging Department
San Francisco, CA
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Re: [Histonet] Help: IHC service

[Colleen Forster via Histonet]

I'd be very interested in this as well...could you shaere Karen.


Thanks.


C

On Mon, Nov 2, 2015 at 11:43 AM, Karen Cai via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> HELP:
>
>
>
> Hi,
>
> Is there anybody can provide me the price list/structure of the custom IHC
> services?
>
>
>
> For example, to test 200 tissue slides and get 200 images, how much does it
> cost?
>
>
>
> Thank you very much in advance,
>
>
>
> Have a nice weekend,
>
>
>
> Best Regards,
>
> Karen
>
> k...@prosci-inc.com
>
> ___
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>
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Re: [Histonet] HELP! Need some old fashioned histology advice

[Jay Lundgren]

 I don't know if the slides were subbed or not, but they looked
improvised.  The point is, at some point the sections are going to have to
be mounted for visualisation, right?  Someone or something is going to look
at the section under magnification?

 If you mount them on glass *before* staining, the whole problem of
batching becomes one of improvising a giant stain rack and giant staining
line.  Don't forget, you need giant cover slips, giant slide folders, and
giant postdocs.
I know that lab glassware is custom ordered every day.  Sounds expensive
and time consuming to set up, but it beats fiddling around with free
floating tissue anyday, IMHO.

I don't know what the Ab penetration is going to be like on a 100um
section, because they weren't using IHC at Genentech, they were doing in
vitro DNA hybridization on the slides.  I'm assuming the section thickness
is necessary because the PI wants to follow axonal pathways and see the
patterns of staining?  Sounds like a fun project.  If you need more help
later you can PM me.

   Sincerely,

  Jay A. Lundgren,
M.S., HTL (ASCP)

On Mon, Jan 5, 2015 at 10:38 PM, Mejia, Mary mary.me...@ucsf.edu wrote:

  Hello Patsy,

  Thank you very much for responding!  Yes  of course, I'll be removing
 the celloidin from each section - we normally
 do this on smaller human whole brainstem sections using ether/100% EA 1:1
 - 3x - 3 minutes each with agitation.  The
 latter type of sections are very easy to work with, however these future
 big boys I'll be getting is another thing.

  Our lab is currently alcohol processing a human whole brain  after the
 last 95% EA - it will be embedded in 2% celloidin
  placed inside a very large desiccator under 20 psi pressure.  This part
 like every step will take some period of time,
 I need to test several different IHC methods  hope one will actually work.

  If you have any further ideas or thoughts on this subject - shoot me an
 email.  Thank you again for responding.

  Maria
  --
 *From:* Patsy Ruegg [prueg...@hotmail.com]
 *Sent:* Sunday, January 04, 2015 7:03 PM
 *To:* Jay Lundgren; Maria Mejia
 *Cc:* Histonet@Lists. Edu; Mejia, Mary
 *Subject:* RE: [Histonet] HELP! Need some old fashioned histology advice

   I have done something similar to this but I used tissue that was fixed
 but not processed and embedded, this is called enblock labeling, I
 infiltrated the fixed tissue with the IHC reagents, in a vial/tube, the
 blocking reagents, then the antibody, then the detection reagents and DAB,
 then dehydrated the tissue.  I used vials or tubes on a platform shaker and
 would infiltrate reagents for days, then after it was done I infiltrated
 and embedded the tissue in glycol methacrylate (GMA) so that I could
 section it, it actually worked.  The tissue was already IHc LABeled so all
 I did to the 5 micron sections after they were cut was a hematoxylin
 counterstain, this was mineralized bone so I had to embedd in something
 hard like GMA to section.

 Will you remove the Celloidin before trying to do the IHC staining?  100
 micron sections might be easy to float/handle using a glass pipette for
 transferring.  Sounds like an interesting project, good luck and feel free
 to ask for advise and keep us posted on your progress.

 Cheers,
 Patsy

 Patsy Ruegg, HT(ASCP)QIHC
 Ruegg IHC Consulting
 40864 E Arkansas Ave
 Bennett, CO 80102
 H 303-644-4538
 C 720-281-5406
 prueg...@hotmail.com



  Date: Sun, 4 Jan 2015 11:58:38 -0600
  From: jaylundg...@gmail.com
  To: mbmph...@gmail.com
  Subject: Re: [Histonet] HELP! Need some old fashioned histology advice
  CC: histonet@lists.utsouthwestern.edu; mary.me...@ucsf.edu
 
  I can help with the old fashioned advice:
 
 
  - 1 scant teaspoon simple syrup
  - 2 dashes Angostura Bitters, plus more to taste
  - 1 half dollar–sized slice orange peel, including pith
  - 2 ounces good-quality rye or bourbon
  - 1 maraschino cherry
 
  As for the Histology, is there any reason you cannot mount the
  sections onto glass slides? When I was working at Genentech they were
  cutting frozen sections through whole rabbits and mounting the sections
 on
  (giant) glass slides. I think that rolling the tissue up, inserting it,
  and then removing it from a glass tube would destroy the tissue.
 
  Sincerely,
 
  Jay A. Lundgren, M.S., HTL
  (ASCP)
 
  On Sat, Jan 3, 2015 at 8:01 PM, Maria Mejia mbmph...@gmail.com wrote:
 
   First, the very best of holidays to everyone.
  
   Now for the histology part. Our lab's focus is on the early stages of
   Alzheimer's Disease in the Brainstem
   using celloidin processing  embedding for IHC staining. This year, our
   lab will be receiving 6 post-mortem
   whole human brains (1 every other month). After fixation, processing 
   celloidin embedding, the whole brain
   will be serially cut at 100um thick. Each brain section will be 5

RE: [Histonet] HELP! Need some old fashioned histology advice

[Mejia, Mary]

Hello Patsy,

Thank you very much for responding!  Yes  of course, I'll be removing the 
celloidin from each section - we normally
do this on smaller human whole brainstem sections using ether/100% EA 1:1 - 3x 
- 3 minutes each with agitation.  The
latter type of sections are very easy to work with, however these future big 
boys I'll be getting is another thing.

Our lab is currently alcohol processing a human whole brain  after the last 
95% EA - it will be embedded in 2% celloidin
 placed inside a very large desiccator under 20 psi pressure.  This part like 
every step will take some period of time,
I need to test several different IHC methods  hope one will actually work.

If you have any further ideas or thoughts on this subject - shoot me an email.  
Thank you again for responding.

Maria

From: Patsy Ruegg [prueg...@hotmail.com]
Sent: Sunday, January 04, 2015 7:03 PM
To: Jay Lundgren; Maria Mejia
Cc: Histonet@Lists. Edu; Mejia, Mary
Subject: RE: [Histonet] HELP! Need some old fashioned histology advice

I have done something similar to this but I used tissue that was fixed but not 
processed and embedded, this is called enblock labeling, I infiltrated the 
fixed tissue with the IHC reagents, in a vial/tube, the blocking reagents, then 
the antibody, then the detection reagents and DAB, then dehydrated the tissue.  
I used vials or tubes on a platform shaker and would infiltrate reagents for 
days, then after it was done I infiltrated and embedded the tissue in glycol 
methacrylate (GMA) so that I could section it, it actually worked.  The tissue 
was already IHc LABeled so all I did to the 5 micron sections after they were 
cut was a hematoxylin counterstain, this was mineralized bone so I had to 
embedd in something hard like GMA to section.

Will you remove the Celloidin before trying to do the IHC staining?  100 micron 
sections might be easy to float/handle using a glass pipette for transferring.  
Sounds like an interesting project, good luck and feel free to ask for advise 
and keep us posted on your progress.

Cheers,
Patsy

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting
40864 E Arkansas Ave
Bennett, CO 80102
H 303-644-4538
C 720-281-5406
prueg...@hotmail.com



 Date: Sun, 4 Jan 2015 11:58:38 -0600
 From: jaylundg...@gmail.com
 To: mbmph...@gmail.com
 Subject: Re: [Histonet] HELP! Need some old fashioned histology advice
 CC: histonet@lists.utsouthwestern.edu; mary.me...@ucsf.edu

 I can help with the old fashioned advice:


 - 1 scant teaspoon simple syrup
 - 2 dashes Angostura Bitters, plus more to taste
 - 1 half dollar–sized slice orange peel, including pith
 - 2 ounces good-quality rye or bourbon
 - 1 maraschino cherry

 As for the Histology, is there any reason you cannot mount the
 sections onto glass slides? When I was working at Genentech they were
 cutting frozen sections through whole rabbits and mounting the sections on
 (giant) glass slides. I think that rolling the tissue up, inserting it,
 and then removing it from a glass tube would destroy the tissue.

 Sincerely,

 Jay A. Lundgren, M.S., HTL
 (ASCP)

 On Sat, Jan 3, 2015 at 8:01 PM, Maria Mejia mbmph...@gmail.com wrote:

  First, the very best of holidays to everyone.
 
  Now for the histology part. Our lab's focus is on the early stages of
  Alzheimer's Disease in the Brainstem
  using celloidin processing  embedding for IHC staining. This year, our
  lab will be receiving 6 post-mortem
  whole human brains (1 every other month). After fixation, processing 
  celloidin embedding, the whole brain
  will be serially cut at 100um thick. Each brain section will be 5 inches
  x 4.5 inches in size.
 
  I will given 250 of these whole brain sections to stain for tau
  IHC...that's 1500 whole brain sections/year!!!
  1) Does anyone have experience doing manual IHC staining of large
  free-floating brain sections?
  2) What type of staining tools, dishes or other essential equipment can
  anyone recommend?
  3) What's the most efficient way to stain 250 sections for batch IHC
  staining - such as transferring batch
  sections (maybe 5-10) from reagent to reagent?
  4) What type of batch apparatus to use?
 
  As for the antibody  ABC steps, I was thinking of placing each section
  inside a large glass cigar tube
  (yep, people use large glass tubes with fitted cap to store cigars), with
  5ml of antibody or ABC reagent  gently agitate on
  a shaker/rotator at room temp during the incubation. Does anyone have
  ideas on this?
 
  Please, any ideas, suggestions or recommendation anyone can provide will
  be most greatly appreciated.
 
  Best regards
  Maria Mejia
  UCSF
  Department of Neurology
  San Francisco, CA
 
 
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 http

Re: [Histonet] HELP! Need some old fashioned histology advice

[Jay Lundgren]

I can help with the old fashioned advice:


   - 1 scant teaspoon simple syrup
   - 2 dashes Angostura Bitters, plus more to taste
   - 1 half dollar–sized slice orange peel, including pith
   - 2 ounces good-quality rye or bourbon
   - 1 maraschino cherry

 As for the Histology, is there any reason you cannot mount the
sections onto glass slides?  When I was working at Genentech they were
cutting frozen sections through whole rabbits and mounting the sections on
(giant) glass slides.  I think that rolling the tissue up, inserting it,
and then removing it from a glass tube would destroy the tissue.

  Sincerely,

Jay A. Lundgren, M.S., HTL
(ASCP)

On Sat, Jan 3, 2015 at 8:01 PM, Maria Mejia mbmph...@gmail.com wrote:

 First, the very best of holidays to everyone.

 Now for the histology part.   Our lab's focus is on the early stages of
 Alzheimer's Disease in the Brainstem
 using celloidin processing  embedding for IHC staining.  This year, our
 lab will be receiving 6 post-mortem
 whole human brains (1 every other month).  After fixation, processing 
 celloidin embedding, the whole brain
 will be serially cut at 100um thick.  Each brain section will be 5 inches
 x 4.5 inches in size.

 I will given 250 of these whole brain sections to stain for tau
 IHC...that's 1500 whole brain sections/year!!!
 1) Does anyone have experience doing manual IHC staining of large
 free-floating brain sections?
 2) What type of staining tools, dishes or other essential equipment can
 anyone recommend?
 3) What's the most efficient way to stain 250 sections for batch IHC
 staining - such as transferring batch
 sections (maybe 5-10) from reagent to reagent?
 4) What type of batch apparatus to use?

 As for the antibody  ABC steps, I was thinking of placing each section
 inside a large glass cigar tube
 (yep, people use large glass tubes with fitted cap to store cigars), with
 5ml of antibody or ABC reagent  gently agitate on
 a shaker/rotator at room temp during the incubation.  Does anyone have
 ideas on this?

 Please, any ideas, suggestions or recommendation anyone can provide will
 be most greatly appreciated.

 Best regards
 Maria Mejia
 UCSF
 Department of Neurology
 San Francisco, CA


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RE: [Histonet] HELP! Need some old fashioned histology advice

[Patsy Ruegg]

I have done something similar to this but I used tissue that was fixed but not 
processed and embedded, this is called enblock labeling, I infiltrated the 
fixed tissue with the IHC reagents, in a vial/tube, the blocking reagents, then 
the antibody, then the detection reagents and DAB, then dehydrated the tissue.  
I used vials or tubes on a platform shaker and would infiltrate reagents for 
days, then after it was done I infiltrated and embedded the tissue in glycol 
methacrylate (GMA) so that I could section it, it actually worked.  The tissue 
was already IHc LABeled so all I did to the 5 micron sections after they were 
cut was a hematoxylin counterstain, this was mineralized bone so I had to 
embedd in something hard like GMA to section.

Will you remove the Celloidin before trying to do the IHC staining?  100 micron 
sections might be easy to float/handle using a glass pipette for transferring.  
Sounds like an interesting project, good luck and feel free to ask for advise 
and keep us posted on your progress.

Cheers,
Patsy

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting
40864 E Arkansas Ave
Bennett, CO 80102
H 303-644-4538
C 720-281-5406
prueg...@hotmail.com



 Date: Sun, 4 Jan 2015 11:58:38 -0600
 From: jaylundg...@gmail.com
 To: mbmph...@gmail.com
 Subject: Re: [Histonet] HELP! Need some old fashioned histology advice
 CC: histonet@lists.utsouthwestern.edu; mary.me...@ucsf.edu
 
 I can help with the old fashioned advice:
 
 
- 1 scant teaspoon simple syrup
- 2 dashes Angostura Bitters, plus more to taste
- 1 half dollar–sized slice orange peel, including pith
- 2 ounces good-quality rye or bourbon
- 1 maraschino cherry
 
  As for the Histology, is there any reason you cannot mount the
 sections onto glass slides?  When I was working at Genentech they were
 cutting frozen sections through whole rabbits and mounting the sections on
 (giant) glass slides.  I think that rolling the tissue up, inserting it,
 and then removing it from a glass tube would destroy the tissue.
 
   Sincerely,
 
 Jay A. Lundgren, M.S., HTL
 (ASCP)
 
 On Sat, Jan 3, 2015 at 8:01 PM, Maria Mejia mbmph...@gmail.com wrote:
 
  First, the very best of holidays to everyone.
 
  Now for the histology part.   Our lab's focus is on the early stages of
  Alzheimer's Disease in the Brainstem
  using celloidin processing  embedding for IHC staining.  This year, our
  lab will be receiving 6 post-mortem
  whole human brains (1 every other month).  After fixation, processing 
  celloidin embedding, the whole brain
  will be serially cut at 100um thick.  Each brain section will be 5 inches
  x 4.5 inches in size.
 
  I will given 250 of these whole brain sections to stain for tau
  IHC...that's 1500 whole brain sections/year!!!
  1) Does anyone have experience doing manual IHC staining of large
  free-floating brain sections?
  2) What type of staining tools, dishes or other essential equipment can
  anyone recommend?
  3) What's the most efficient way to stain 250 sections for batch IHC
  staining - such as transferring batch
  sections (maybe 5-10) from reagent to reagent?
  4) What type of batch apparatus to use?
 
  As for the antibody  ABC steps, I was thinking of placing each section
  inside a large glass cigar tube
  (yep, people use large glass tubes with fitted cap to store cigars), with
  5ml of antibody or ABC reagent  gently agitate on
  a shaker/rotator at room temp during the incubation.  Does anyone have
  ideas on this?
 
  Please, any ideas, suggestions or recommendation anyone can provide will
  be most greatly appreciated.
 
  Best regards
  Maria Mejia
  UCSF
  Department of Neurology
  San Francisco, CA
 
 
  ___
  Histonet mailing list
  Histonet@lists.utsouthwestern.edu
  http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 
 ___
 Histonet mailing list
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 http://lists.utsouthwestern.edu/mailman/listinfo/histonet
  
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RE: [Histonet] HELP

[Elizabeth Chlipala]

Hans

We use the antibody from Serotec, it works quite nicely.  MCA711, proteinase K 
digestion,  rabbit anti-rat secondary and then Envision Rabbit (polymer).  We 
use this antibody at a 1:1200 dilution.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
l...@premierlab.com
www.premierlab.com

March 10, 2014 is Histotechnology Professionals Day

Ship to Address:

Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Hans B Snyder
Sent: Thursday, November 06, 2014 10:31 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] HELP

Hello All,

Does anyone have a protocol for the Anti-Neutrophil antibody [NIMP-R14]
(ab2557) for mouse tissue?  I have been trying to extinguish the background 
staining but cannot find the right combination.
​ We are doing this by hand. ​
 The recommended dilution for this antibody is 1:50 with HIER.   See used
protocol below.

So far I have tried:
1. concentrations 1:25, 1:50, 1:75, 1:100  - 1:50 gives very high background 
but good positive staining, 1:100 gives low background but not much positive 
staining.

2. HIER - 3, 5, 10,  20, minutes  @ 96C, 20 minutes at 60C  all give the same 
results - too much back ground

3. We have tried using the H2O2 before the primary and after, increasing the 
time from 30 minutes to 45 minutes.

4. Tried incubating in the primary for shorter time (20 minutes) and longer 
overnight at 4C.

5. Tried blocking using 3 different serums, first each one then combinations 
and longer times.

6. Tried cutting the DAB concentration in 1/2 then 1/3.


We have not tried enzyme or acid epitope retrieval yet.



*Does anyone have a working protocol or suggestions on what to try?*


​Thank you in advance.​





Procedure:

Deparaffinize

1.  Heat slides to 60C for 10 minutes (oven).

2.  Dry slides at room temp 5 minutes.

3.  Xylene 5 minutes.

4.  Xylene 5 minutes.

5.  100% ethanol 5 minutes (dehydration).

6.  100% ethanol 2 minutes (dehydration).

7.  95% ethanol for 2 minutes (rehydration).

8.  70% ethanol for 2 minutes (rehydration).

9.   Run in DI water for 10 minutes.



Antigen Retrieval

1.  Submerge in 0.01M Citrate Buffer (pH 6.0) at 96C for 5 minutes.

2.  Cold running water 5 minutes.



Staining

1.  Wash in PBS for 1 min.

2.  Permeabilize with 0.1% Triton X / 2.5% horse serum + 2.5% fetal
bovine serum for 2 hours.

3.  Incubate in primary antibody 30-45 minutes at in 2.5% horse serum
in PBS.

4.  Wash 3x in PBS/tween for 5 minutes each.

5.  Fresh 3.0% hydrogen peroxide (H2O2) 2x for 20 minutes each.

6.  Wash 3x PBS 2 minutes each.

7.  Incubate with anti-Rat Ig impress solution (according to vector’s
instructions) 45 minutes.

8.  Wash 3x in PBS for 5 minutes each.

9.  Prepare Impact DAB substrate (see insert).

10.  Using a light microscope, incubate sections with 50-100ul, until desired 
darkness occurs (5-30 seconds).

11.  Stop DAB reaction using deionized water.

12.  Wash in DH2O for 5 minutes

13.  Counterstain in Harris hematoxylin for 30 seconds.

14.  Wash in DH2O for 5 minutes.

15.  Dehydrate 2x in 95% alcohol for 30 seconds each.

16.  Dehydrate 3x in 100% alcohol for 2 minutes each.

17.  Clear 2x in xylene 5 minutes each.

18.  Mount coverslips using Leica mounting media.


Hans B Snyder
Histologistics
60 Prescott Street
Worcester, MA 01605
508-308-7800
h...@histologistics.com
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RE: [Histonet] HELP

[Connolly, Brett M]

We also use the MCA711 antibody (at 1.0 ug/mL, 1 hr.), but with HIER in citrate 
buffer using a Biocare Decloaker pressure cooker and Biocare's rat-on mouse HRP 
polymer detection. We  get very nice staining with no background. I was never 
too impressed with ImPRESS.

Here are the basics :
- Deparaffinize and hydrate to dH20
- Perform HIER, cool for 20 min then wash in H20
- 3.0% H2O2 - 20 min
- Serum free block (Biocare Sniper) 30 min
- Incubate with MCA771 1 hr.
- Incubate with Biocare rat-on-Mouse kit (per instructions)
- DAB - 5 min.
 Washes are with PBS/0.1% Tween

Brett


Brett M. Connolly, Ph.D.
Principle Scientist, Imaging Dept.
Merck  Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
brett_conno...@merck.com
T- 215-652-2501
F- 215-993-6803




-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth 
Chlipala
Sent: Thursday, November 06, 2014 12:41 PM
To: Hans B Snyder; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] HELP

Hans

We use the antibody from Serotec, it works quite nicely.  MCA711, proteinase K 
digestion,  rabbit anti-rat secondary and then Envision Rabbit (polymer).  We 
use this antibody at a 1:1200 dilution.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
l...@premierlab.com
www.premierlab.com

March 10, 2014 is Histotechnology Professionals Day

Ship to Address:

Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Hans B Snyder
Sent: Thursday, November 06, 2014 10:31 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] HELP

Hello All,

Does anyone have a protocol for the Anti-Neutrophil antibody [NIMP-R14]
(ab2557) for mouse tissue?  I have been trying to extinguish the background 
staining but cannot find the right combination.
​ We are doing this by hand. ​
 The recommended dilution for this antibody is 1:50 with HIER.   See used
protocol below.

So far I have tried:
1. concentrations 1:25, 1:50, 1:75, 1:100  - 1:50 gives very high background 
but good positive staining, 1:100 gives low background but not much positive 
staining.

2. HIER - 3, 5, 10,  20, minutes  @ 96C, 20 minutes at 60C  all give the same 
results - too much back ground

3. We have tried using the H2O2 before the primary and after, increasing the 
time from 30 minutes to 45 minutes.

4. Tried incubating in the primary for shorter time (20 minutes) and longer 
overnight at 4C.

5. Tried blocking using 3 different serums, first each one then combinations 
and longer times.

6. Tried cutting the DAB concentration in 1/2 then 1/3.


We have not tried enzyme or acid epitope retrieval yet.



*Does anyone have a working protocol or suggestions on what to try?*


​Thank you in advance.​





Procedure:

Deparaffinize

1.  Heat slides to 60C for 10 minutes (oven).

2.  Dry slides at room temp 5 minutes.

3.  Xylene 5 minutes.

4.  Xylene 5 minutes.

5.  100% ethanol 5 minutes (dehydration).

6.  100% ethanol 2 minutes (dehydration).

7.  95% ethanol for 2 minutes (rehydration).

8.  70% ethanol for 2 minutes (rehydration).

9.   Run in DI water for 10 minutes.



Antigen Retrieval

1.  Submerge in 0.01M Citrate Buffer (pH 6.0) at 96C for 5 minutes.

2.  Cold running water 5 minutes.



Staining

1.  Wash in PBS for 1 min.

2.  Permeabilize with 0.1% Triton X / 2.5% horse serum + 2.5% fetal
bovine serum for 2 hours.

3.  Incubate in primary antibody 30-45 minutes at in 2.5% horse serum
in PBS.

4.  Wash 3x in PBS/tween for 5 minutes each.

5.  Fresh 3.0% hydrogen peroxide (H2O2) 2x for 20 minutes each.

6.  Wash 3x PBS 2 minutes each.

7.  Incubate with anti-Rat Ig impress solution (according to vector’s
instructions) 45 minutes.

8.  Wash 3x in PBS for 5 minutes each.

9.  Prepare Impact DAB substrate (see insert).

10.  Using a light microscope, incubate sections with 50-100ul, until desired 
darkness occurs (5-30 seconds).

11.  Stop DAB reaction using deionized water.

12.  Wash in DH2O for 5 minutes

13.  Counterstain in Harris hematoxylin for 30 seconds.

14.  Wash in DH2O for 5 minutes.

15.  Dehydrate 2x in 95% alcohol for 30 seconds each.

16.  Dehydrate 3x in 100% alcohol for 2 minutes each.

17.  Clear 2x in xylene 5 minutes each.

18.  Mount coverslips using Leica mounting media.


Hans B Snyder
Histologistics
60 Prescott Street
Worcester, MA 01605
508-308-7800
h...@histologistics.com
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Re: [Histonet] HELP- Cryosectioning FAT!

[Balasubbramanian, Dakshnapriya]

Thanks Sandra! My next step was to try that.

Dakshna

- Original Message -
From: Sandra E. Esparza sespa...@seton.org
To: Dakshnapriya Balasubbramanian dakshnapr...@neo.tamu.edu
Sent: Tuesday, May 20, 2014 10:34:25 AM
Subject: RE: [Histonet] HELP- Cryosectioning FAT!

You might want to dry Liquid Nitrogen on the face of the block before you cut 
it.  

Sandra

Sandra Esparza HT (ASCP), QIHC
Histotechnologist Mohs
Austin Dermatologic Surgery Center
512-324-7468  x84027
sespa...@seton.org


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
Balasubbramanian, Dakshnapriya
Sent: Tuesday, May 20, 2014 10:23 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] HELP- Cryosectioning FAT!

Hi all,

I've been having problems with cryosectioning fatty tissue for a long time now. 
The section leaves a hole in the middle. I know this is probably a much 
discussed topic, but I've tried a lot of strategies with no luck. The tissue is 
of mouse origin and has micro-lesions buried inside fat. So the fat needs to be 
cut in order to get to the lesion. I started with a temperature of -17C and 
tried upto -25C and also at -50C; didn't work at any of these temperatures. I 
tried rubbing the block, blade and anti-roll plate with dry ice-again,no luck. 

I rapid-freeze the tissue with OCT in LN2 and then store the block at -80C. 
Could I try sectioning the block directly from -80C? (never done that). 

Any other suggestions on how to tackle this?

Any advice is much appreciated!!

Thanks,
Dakshna

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Re: [Histonet] HELP- Cryosectioning FAT!

[Balasubbramanian, Dakshnapriya]

Hi Laurie,

No, the tissue hasn't been fixed before freezing. My prof doesn't prefer the 
method, but if nothing else works, we might give it a try. 

Any suggested protocols for fixation?

Thanks!
Dakshna

- Original Message -
From: Laurie J King king.lau...@marshfieldclinic.org
To: Dakshnapriya Balasubbramanian dakshnapr...@neo.tamu.edu
Sent: Tuesday, May 20, 2014 10:37:35 AM
Subject: RE: [Histonet] HELP- Cryosectioning FAT!

Dakshna,

Has this tissue been fixed before freezing by any chance?

laurie

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of 
Balasubbramanian, Dakshnapriya
Sent: Tuesday, May 20, 2014 10:23 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] HELP- Cryosectioning FAT!

Hi all,

I've been having problems with cryosectioning fatty tissue for a long time now. 
The section leaves a hole in the middle. I know this is probably a much 
discussed topic, but I've tried a lot of strategies with no luck. The tissue is 
of mouse origin and has micro-lesions buried inside fat. So the fat needs to be 
cut in order to get to the lesion. I started with a temperature of -17C and 
tried upto -25C and also at -50C; didn't work at any of these temperatures. I 
tried rubbing the block, blade and anti-roll plate with dry ice-again,no luck. 

I rapid-freeze the tissue with OCT in LN2 and then store the block at -80C. 
Could I try sectioning the block directly from -80C? (never done that). 

Any other suggestions on how to tackle this?

Any advice is much appreciated!!

Thanks,
Dakshna

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Re: [Histonet] Help for identifying the blue stained structure

[Marvin Hanna]

Hi Rui,

You uploaded a tif image and a number of browsers don't support tif 
images. Jpeg, gif and png images are the best image formats to use 
because they are universally supported. I converted your image to a jpeg 
image and posted it at:


http://histosearch.com/imageupload/help-for-identifying-the-blue-stained-structure-jpeg/

Best Regards,

Marvin Hanna


On 05/17/2014 06:39 PM, Rui TAHARA wrote:


Hello,
I have an embryonic sample that
decalcified, paraffin embedded, and stained with Mallory Trichrome (Aniline
Blue, Orange G, Acid Fuchsin). I will upload the image in the Histonet Images. 
This
is a cranial region where the bone is being resorbed, so I expected to see the
bone (dark blue in trabecular), and red blood, and adjacent white spaces that
is being resorbed. Instead, there is a very uniform, granular structure stained
with blue without any nuclei at the resorbed regions. This structure looks like
almost crystal or some kind of secretion leakage from the ossifying bone. I
need a help to identify this blue stained things. I don't think this is osteoid,
because at later stage embryos, there is no bone at this region.

If you have some suggestions for other
stain to identify this blue thing, or help identifying this, I really
appreciate it.

  


Thank you in advance,

  


Rui




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RE: [Histonet] Help for identifying the blue stained structure

[Rui TAHARA]

Hi, 

Thank you so much. 
I was wondering why the image has not been seen on the site, and thought its 
been processed. 

Thank you for uploading the image. 

Rui 

Date: Sun, 18 May 2014 17:47:21 -0400
From: mha...@histosearch.com
To: ru...@hotmail.com; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Help for identifying the blue stained structure


  

  
  
Hi Rui,



You uploaded a tif image and a number of browsers don't support tif
images. Jpeg, gif and png images are the best image formats to use
because they are universally supported. I converted your image to a
jpeg image and posted it at:




http://histosearch.com/imageupload/help-for-identifying-the-blue-stained-structure-jpeg/



Best Regards,



Marvin Hanna





On 05/17/2014 06:39 PM, Rui TAHARA
  wrote:



  Hello, 
I have an embryonic sample that
decalcified, paraffin embedded, and stained with Mallory Trichrome (Aniline
Blue, Orange G, Acid Fuchsin). I will upload the image in the Histonet Images. 
This
is a cranial region where the bone is being resorbed, so I expected to see the
bone (dark blue in trabecular), and red blood, and adjacent white spaces that
is being resorbed. Instead, there is a very uniform, granular structure stained
with blue without any nuclei at the resorbed regions. This structure looks like
almost crystal or some kind of secretion leakage from the ossifying bone. I
need a help to identify this blue stained things. I don’t think this is osteoid,
because at later stage embryos, there is no bone at this region. 

If you have some suggestions for other
stain to identify this blue thing, or help identifying this, I really
appreciate it. 

 

Thank you in advance, 

 

Rui 

  
  

  
  

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Re: [Histonet] Help on get PDF file paper from J. Histotechnology

[Dorothy Hu]

Thanks very much Jean. I will call NSH office. Dorothy


On Fri, Jan 17, 2014 at 2:53 PM, Mitchell Jean A jmitch...@uwhealth.orgwrote:

 Dorothy:  I know that there have been some issues with Maney Online
 lately.  I suggest your contact the NSH office and they should be able to
 assist you.  I had the same problem last week and it was handled very
 quickly.


 Jean Mitchell, BS HT (ASCP)
 University of Wisconsin Hospital  Clinics
 Neuromuscular Laboratory Manager
 600 Highland Avenue
 Madison, WI  53792-5132


 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Dorothy Hu
 Sent: Friday, January 17, 2014 8:57 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Help on get PDF file paper from J. Histotechnology

 Hi Histonetters,

 Would you please let me know why I can not get PDF file from J
 Histotechnology any more?

 I renewed my membership after new year and changed my password since i
 don't remember  my old one. I can get in Maney Online for only Abstract,
 but not whole article.

 The paper I tried to get is:
 Parallel experience of two different lab with initiator perkadox 16 for
 polymerization of MMA.
 Vol 17, No 4, pp.343-348.

 Thanks in advance.

 Dorothy
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Re: [Histonet] Help

[Emily Sours]

FACEPALM.JPG

this is not even worth an actual jpg.
sigh.

By bitching and bitching and bitching, they could exhaust the drama of
their own horror stories. Grow bored. Only then could they accept a new
story for their lives. Move forward.

-Chuck Palahniuk, Haunted


On Wed, Sep 11, 2013 at 7:51 PM, Jay Lundgren jaylundg...@gmail.com wrote:

 lol


 On Wed, Sep 11, 2013 at 5:56 PM, Bain,Virginia veb...@mdanderson.org
 wrote:

  Googling 'unsubscribe histonet' points me to this link:
  http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 
  If you scroll to the bottom of the page you will find a box where you can
  type your e-mail address to unsubscribe or edit options.  Have you tried
  that?
 
  Cheers.
 
  --
  Virginia Bain
  Postdoctoral Fellow
  Richie Lab
  512-237-6443
 
 
 
 
 
  On 9/11/13 5:38 PM, nancy mcvay njmcvay...@gmail.com wrote:
 
  Get me off your mailing list.
  I  have tried and keep getting these things.
  Over and over you repeat the same questions and answers ,   over and
 over,
  over  and over,  over and over,  over and over.
  
  Do  you get the message?
  Do you get the message?
  Do you get the message?
  Get me off your mailing list
  Get me off your mailing list
  
  
  Today  I received four different email  tirades from you.
  
  
  Please get me off your mailing list.
  
  I am to the point of reporting it as spam.
  
  njmcvay...@gmail.com
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Re: [Histonet] Help

[Bain,Virginia]

Googling 'unsubscribe histonet' points me to this link:
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

If you scroll to the bottom of the page you will find a box where you can
type your e-mail address to unsubscribe or edit options.  Have you tried
that?

Cheers.

-- 
Virginia Bain
Postdoctoral Fellow
Richie Lab
512-237-6443





On 9/11/13 5:38 PM, nancy mcvay njmcvay...@gmail.com wrote:

Get me off your mailing list.
I  have tried and keep getting these things.
Over and over you repeat the same questions and answers ,   over and over,
over  and over,  over and over,  over and over.

Do  you get the message?
Do you get the message?
Do you get the message?
Get me off your mailing list
Get me off your mailing list


Today  I received four different email  tirades from you.


Please get me off your mailing list.

I am to the point of reporting it as spam.

njmcvay...@gmail.com
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Re: [Histonet] Help

[Jay Lundgren]

lol


On Wed, Sep 11, 2013 at 5:56 PM, Bain,Virginia veb...@mdanderson.orgwrote:

 Googling 'unsubscribe histonet' points me to this link:
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet

 If you scroll to the bottom of the page you will find a box where you can
 type your e-mail address to unsubscribe or edit options.  Have you tried
 that?

 Cheers.

 --
 Virginia Bain
 Postdoctoral Fellow
 Richie Lab
 512-237-6443





 On 9/11/13 5:38 PM, nancy mcvay njmcvay...@gmail.com wrote:

 Get me off your mailing list.
 I  have tried and keep getting these things.
 Over and over you repeat the same questions and answers ,   over and over,
 over  and over,  over and over,  over and over.
 
 Do  you get the message?
 Do you get the message?
 Do you get the message?
 Get me off your mailing list
 Get me off your mailing list
 
 
 Today  I received four different email  tirades from you.
 
 
 Please get me off your mailing list.
 
 I am to the point of reporting it as spam.
 
 njmcvay...@gmail.com
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RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

[Weems, Joyce K.]

Yes, it is for review of material for molecular testing and can be added at any 
time as long as it is not ordered at the time the case is in process. If it is 
more than 30 days after the patient has been discharged, it is considered 
archived (according to Medicare) and we register it for a new acct number and 
bill it alone. Otherwise it is a late bill on current visit.

We do this regardless if the patient is Medicare or not - just for consistency. 
I am not aware of any problems.

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's 
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contain information that is privileged and confidential.  Any unauthorized 
review, use, disclosure, or distribution is prohibited. If you are not the 
intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Natalie Nagy
Sent: Thursday, January 10, 2013 10:36 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Help with CPT code 88363 for archived tissue retrieval

Hi everyone,
   I just have a question about CPT code 88363, first can it be 
used for pulling blocks for Oncotype DX testing, also is there a time limit on 
when this code can be used? Does it have to be within a year, a month, etc...of 
when the patient account went active?

Thanks for all the help,

Natalie J. Nagy (HT)ASCP
Histology Supervisor
Holyoke Medical Center


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Re: [Histonet] Help with CPT code 88363 for archived tissue retrieval

[Mark Tarango]

It can't be used to just pull blocks.  The slides have to be reviewed and
the best block chosen by a pathologist.  If there is only one block then
the pathologist needs to look at the slides and determine if there is
enough tissue for molecular testing.  It's a professional charge.

Use it on re-accessioned cases for which molecular testing is requested and
the best block needs to be chosen.

Yes we use it when sending out for Oncotype DX if the case was signed out
over 30 days before.

Mark

On Thu, Jan 10, 2013 at 7:36 AM, Natalie Nagy 
nagy_nata...@holyokehealth.com wrote:

 Hi everyone,
I just have a question about CPT code 88363, first can
 it be used for pulling blocks for Oncotype DX testing, also is there a time
 limit on when this code can be used? Does it have to be within a year, a
 month, etc...of when the patient account went active?

 Thanks for all the help,

 Natalie J. Nagy (HT)ASCP
 Histology Supervisor
 Holyoke Medical Center


 CONFIDENTIALITY NOTICE: This email communication and any attachments may
 contain confidential and privileged information for the use of the
 designated recipients named above. If you are not the intended recipient,
 you are hereby notified that you have received this communication in error
 and that any review, disclosure, dissemination, distribution or copying of
 it or its contents is prohibited. If you have received this communication
 in error, please reply to the sender immediately and destroy all copies of
 this communication and any attachments. For further information regarding
 Holyoke Medical Center's privacy policy, Please visit our Internet web site
 at http://www.holyokehealth.com
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

[Weems, Joyce K.]

And I should explain the reason I most know this.. our pathologists were denied 
because the tech charge hadn't been entered yet. So now I make sure the tech 
charge is entered before sending to the pathologists billing folks.

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's 
Hospital and is intended for the sole use of the intended recipient(s).  It may 
contain information that is privileged and confidential.  Any unauthorized 
review, use, disclosure, or distribution is prohibited. If you are not the 
intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email.


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark Tarango
Sent: Thursday, January 10, 2013 2:01 PM
To: Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Help with CPT code 88363 for archived tissue retrieval

It can't be used to just pull blocks.  The slides have to be reviewed and the 
best block chosen by a pathologist.  If there is only one block then the 
pathologist needs to look at the slides and determine if there is enough tissue 
for molecular testing.  It's a professional charge.

Use it on re-accessioned cases for which molecular testing is requested and the 
best block needs to be chosen.

Yes we use it when sending out for Oncotype DX if the case was signed out over 
30 days before.

Mark

On Thu, Jan 10, 2013 at 7:36 AM, Natalie Nagy  nagy_nata...@holyokehealth.com 
wrote:

 Hi everyone,
I just have a question about CPT code 88363, first
 can it be used for pulling blocks for Oncotype DX testing, also is
 there a time limit on when this code can be used? Does it have to be
 within a year, a month, etc...of when the patient account went active?

 Thanks for all the help,

 Natalie J. Nagy (HT)ASCP
 Histology Supervisor
 Holyoke Medical Center


 CONFIDENTIALITY NOTICE: This email communication and any attachments
 may contain confidential and privileged information for the use of the
 designated recipients named above. If you are not the intended
 recipient, you are hereby notified that you have received this
 communication in error and that any review, disclosure, dissemination,
 distribution or copying of it or its contents is prohibited. If you
 have received this communication in error, please reply to the sender
 immediately and destroy all copies of this communication and any
 attachments. For further information regarding Holyoke Medical
 Center's privacy policy, Please visit our Internet web site at
 http://www.holyokehealth.com
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

[Weems, Joyce K.]

It is also a technical charge, as well.

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's 
Hospital and is intended for the sole use of the intended recipient(s).  It may 
contain information that is privileged and confidential.  Any unauthorized 
review, use, disclosure, or distribution is prohibited. If you are not the 
intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email.


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark Tarango
Sent: Thursday, January 10, 2013 2:01 PM
To: Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Help with CPT code 88363 for archived tissue retrieval

It can't be used to just pull blocks.  The slides have to be reviewed and the 
best block chosen by a pathologist.  If there is only one block then the 
pathologist needs to look at the slides and determine if there is enough tissue 
for molecular testing.  It's a professional charge.

Use it on re-accessioned cases for which molecular testing is requested and the 
best block needs to be chosen.

Yes we use it when sending out for Oncotype DX if the case was signed out over 
30 days before.

Mark

On Thu, Jan 10, 2013 at 7:36 AM, Natalie Nagy  nagy_nata...@holyokehealth.com 
wrote:

 Hi everyone,
I just have a question about CPT code 88363, first
 can it be used for pulling blocks for Oncotype DX testing, also is
 there a time limit on when this code can be used? Does it have to be
 within a year, a month, etc...of when the patient account went active?

 Thanks for all the help,

 Natalie J. Nagy (HT)ASCP
 Histology Supervisor
 Holyoke Medical Center


 CONFIDENTIALITY NOTICE: This email communication and any attachments
 may contain confidential and privileged information for the use of the
 designated recipients named above. If you are not the intended
 recipient, you are hereby notified that you have received this
 communication in error and that any review, disclosure, dissemination,
 distribution or copying of it or its contents is prohibited. If you
 have received this communication in error, please reply to the sender
 immediately and destroy all copies of this communication and any
 attachments. For further information regarding Holyoke Medical
 Center's privacy policy, Please visit our Internet web site at
 http://www.holyokehealth.com
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

[Mike Pence]

So are you putting the charge thru twice or is the charge for the 88363
divided out into tech and prof.?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Weems,
Joyce K.
Sent: Thursday, January 10, 2013 1:06 PM
To: 'Mark Tarango'; Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help with CPT code 88363 for archived tissue
retrieval


And I should explain the reason I most know this.. our pathologists were
denied because the tech charge hadn't been entered yet. So now I make
sure the tech charge is entered before sending to the pathologists
billing folks.

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's
Hospital and is intended for the sole use of the intended recipient(s).
It may contain information that is privileged and confidential.  Any
unauthorized review, use, disclosure, or distribution is prohibited. If
you are not the intended recipient, please delete this message, and
reply to the sender regarding the error in a separate email.


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark
Tarango
Sent: Thursday, January 10, 2013 2:01 PM
To: Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Help with CPT code 88363 for archived tissue
retrieval

It can't be used to just pull blocks.  The slides have to be reviewed
and the best block chosen by a pathologist.  If there is only one block
then the pathologist needs to look at the slides and determine if there
is enough tissue for molecular testing.  It's a professional charge.

Use it on re-accessioned cases for which molecular testing is requested
and the best block needs to be chosen.

Yes we use it when sending out for Oncotype DX if the case was signed
out over 30 days before.

Mark

On Thu, Jan 10, 2013 at 7:36 AM, Natalie Nagy 
nagy_nata...@holyokehealth.com wrote:

 Hi everyone,
I just have a question about CPT code 88363, first 
 can it be used for pulling blocks for Oncotype DX testing, also is 
 there a time limit on when this code can be used? Does it have to be 
 within a year, a month, etc...of when the patient account went active?

 Thanks for all the help,

 Natalie J. Nagy (HT)ASCP
 Histology Supervisor
 Holyoke Medical Center


 CONFIDENTIALITY NOTICE: This email communication and any attachments 
 may contain confidential and privileged information for the use of the

 designated recipients named above. If you are not the intended 
 recipient, you are hereby notified that you have received this 
 communication in error and that any review, disclosure, dissemination,

 distribution or copying of it or its contents is prohibited. If you 
 have received this communication in error, please reply to the sender 
 immediately and destroy all copies of this communication and any 
 attachments. For further information regarding Holyoke Medical 
 Center's privacy policy, Please visit our Internet web site at 
 http://www.holyokehealth.com 
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu 
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

[Weems, Joyce K.]

Divided - but can be billed globally if that is how you bill. The professional 
uses the 26 modifier. 

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's 
Hospital and is intended for the sole use of the intended recipient(s).  It may 
contain information that is privileged and confidential.  Any unauthorized 
review, use, disclosure, or distribution is prohibited. If you are not the 
intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email. 


-Original Message-
From: Mike Pence [mailto:mpe...@grhs.net] 
Sent: Thursday, January 10, 2013 2:28 PM
To: Weems, Joyce K.; Mark Tarango; Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

So are you putting the charge thru twice or is the charge for the 88363 divided 
out into tech and prof.?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce K.
Sent: Thursday, January 10, 2013 1:06 PM
To: 'Mark Tarango'; Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval


And I should explain the reason I most know this.. our pathologists were denied 
because the tech charge hadn't been entered yet. So now I make sure the tech 
charge is entered before sending to the pathologists billing folks.

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's 
Hospital and is intended for the sole use of the intended recipient(s).
It may contain information that is privileged and confidential.  Any 
unauthorized review, use, disclosure, or distribution is prohibited. If you are 
not the intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email.


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark Tarango
Sent: Thursday, January 10, 2013 2:01 PM
To: Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Help with CPT code 88363 for archived tissue retrieval

It can't be used to just pull blocks.  The slides have to be reviewed and the 
best block chosen by a pathologist.  If there is only one block then the 
pathologist needs to look at the slides and determine if there is enough tissue 
for molecular testing.  It's a professional charge.

Use it on re-accessioned cases for which molecular testing is requested and the 
best block needs to be chosen.

Yes we use it when sending out for Oncotype DX if the case was signed out over 
30 days before.

Mark

On Thu, Jan 10, 2013 at 7:36 AM, Natalie Nagy  nagy_nata...@holyokehealth.com 
wrote:

 Hi everyone,
I just have a question about CPT code 88363, first 
 can it be used for pulling blocks for Oncotype DX testing, also is 
 there a time limit on when this code can be used? Does it have to be 
 within a year, a month, etc...of when the patient account went active?

 Thanks for all the help,

 Natalie J. Nagy (HT)ASCP
 Histology Supervisor
 Holyoke Medical Center


 CONFIDENTIALITY NOTICE: This email communication and any attachments 
 may contain confidential and privileged information for the use of the

 designated recipients named above. If you are not the intended 
 recipient, you are hereby notified that you have received this 
 communication in error and that any review, disclosure, dissemination,

 distribution or copying of it or its contents is prohibited. If you 
 have received this communication in error, please reply to the sender 
 immediately and destroy all copies of this communication and any 
 attachments. For further information regarding Holyoke Medical 
 Center's privacy policy, Please visit our Internet web site at 
 http://www.holyokehealth.com 
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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(including any attachments

RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

[Richard Cartun]

I'm not 100% sure, but I don't think this CPT code has a Technical component.

Richard

Richard W. Cartun, MS, PhD
Director, Histology  Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-2204 Fax


 Mike Pence mpe...@grhs.net 1/10/2013 2:27 PM 
So are you putting the charge thru twice or is the charge for the 88363
divided out into tech and prof.?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Weems,
Joyce K.
Sent: Thursday, January 10, 2013 1:06 PM
To: 'Mark Tarango'; Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu 
Subject: RE: [Histonet] Help with CPT code 88363 for archived tissue
retrieval


And I should explain the reason I most know this.. our pathologists were
denied because the tech charge hadn't been entered yet. So now I make
sure the tech charge is entered before sending to the pathologists
billing folks.

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org 



www.saintjosephsatlanta.org 
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's
Hospital and is intended for the sole use of the intended recipient(s).
It may contain information that is privileged and confidential.  Any
unauthorized review, use, disclosure, or distribution is prohibited. If
you are not the intended recipient, please delete this message, and
reply to the sender regarding the error in a separate email.


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark
Tarango
Sent: Thursday, January 10, 2013 2:01 PM
To: Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu 
Subject: Re: [Histonet] Help with CPT code 88363 for archived tissue
retrieval

It can't be used to just pull blocks.  The slides have to be reviewed
and the best block chosen by a pathologist.  If there is only one block
then the pathologist needs to look at the slides and determine if there
is enough tissue for molecular testing.  It's a professional charge.

Use it on re-accessioned cases for which molecular testing is requested
and the best block needs to be chosen.

Yes we use it when sending out for Oncotype DX if the case was signed
out over 30 days before.

Mark

On Thu, Jan 10, 2013 at 7:36 AM, Natalie Nagy 
nagy_nata...@holyokehealth.com wrote:

 Hi everyone,
I just have a question about CPT code 88363, first 
 can it be used for pulling blocks for Oncotype DX testing, also is 
 there a time limit on when this code can be used? Does it have to be 
 within a year, a month, etc...of when the patient account went active?

 Thanks for all the help,

 Natalie J. Nagy (HT)ASCP
 Histology Supervisor
 Holyoke Medical Center


 CONFIDENTIALITY NOTICE: This email communication and any attachments 
 may contain confidential and privileged information for the use of the

 designated recipients named above. If you are not the intended 
 recipient, you are hereby notified that you have received this 
 communication in error and that any review, disclosure, dissemination,

 distribution or copying of it or its contents is prohibited. If you 
 have received this communication in error, please reply to the sender 
 immediately and destroy all copies of this communication and any 
 attachments. For further information regarding Holyoke Medical 
 Center's privacy policy, Please visit our Internet web site at 
 http://www.holyokehealth.com 
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu 
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

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RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

[Bell, Lynne]

I agree, Dr. Cartun.  I believe that the CPT Coding book specifically says that 
it is only a professional charge.

Lynne Bell, HT (ASCP)
Histology Team Leader
Central Vermont Medical Center
130 Fisher Road
Berlin, VT  05641
802-371-4923
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Richard Cartun
Sent: Thursday, January 10, 2013 2:47 PM
To: Joyce K. Weems; Mark Tarango; Mike Pence; Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

I'm not 100% sure, but I don't think this CPT code has a Technical component.

Richard

Richard W. Cartun, MS, PhD
Director, Histology  Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-2204 Fax


 Mike Pence mpe...@grhs.net 1/10/2013 2:27 PM 
So are you putting the charge thru twice or is the charge for the 88363
divided out into tech and prof.?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Weems,
Joyce K.
Sent: Thursday, January 10, 2013 1:06 PM
To: 'Mark Tarango'; Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu 
Subject: RE: [Histonet] Help with CPT code 88363 for archived tissue
retrieval


And I should explain the reason I most know this.. our pathologists were
denied because the tech charge hadn't been entered yet. So now I make
sure the tech charge is entered before sending to the pathologists
billing folks.

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org 



www.saintjosephsatlanta.org 
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's
Hospital and is intended for the sole use of the intended recipient(s).
It may contain information that is privileged and confidential.  Any
unauthorized review, use, disclosure, or distribution is prohibited. If
you are not the intended recipient, please delete this message, and
reply to the sender regarding the error in a separate email.


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark
Tarango
Sent: Thursday, January 10, 2013 2:01 PM
To: Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu 
Subject: Re: [Histonet] Help with CPT code 88363 for archived tissue
retrieval

It can't be used to just pull blocks.  The slides have to be reviewed
and the best block chosen by a pathologist.  If there is only one block
then the pathologist needs to look at the slides and determine if there
is enough tissue for molecular testing.  It's a professional charge.

Use it on re-accessioned cases for which molecular testing is requested
and the best block needs to be chosen.

Yes we use it when sending out for Oncotype DX if the case was signed
out over 30 days before.

Mark

On Thu, Jan 10, 2013 at 7:36 AM, Natalie Nagy 
nagy_nata...@holyokehealth.com wrote:

 Hi everyone,
I just have a question about CPT code 88363, first 
 can it be used for pulling blocks for Oncotype DX testing, also is 
 there a time limit on when this code can be used? Does it have to be 
 within a year, a month, etc...of when the patient account went active?

 Thanks for all the help,

 Natalie J. Nagy (HT)ASCP
 Histology Supervisor
 Holyoke Medical Center


 CONFIDENTIALITY NOTICE: This email communication and any attachments 
 may contain confidential and privileged information for the use of the

 designated recipients named above. If you are not the intended 
 recipient, you are hereby notified that you have received this 
 communication in error and that any review, disclosure, dissemination,

 distribution or copying of it or its contents is prohibited. If you 
 have received this communication in error, please reply to the sender 
 immediately and destroy all copies of this communication and any 
 attachments. For further information regarding Holyoke Medical 
 Center's privacy policy, Please visit our Internet web site at 
 http://www.holyokehealth.com 
 ___
 Histonet mailing list
 Histonet@lists.utsouthwestern.edu 
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

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or copying of this message (including any attachments

RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

[Weems, Joyce K.]

Maybe so, but I am getting reimbursed for tech only. 

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's 
Hospital and is intended for the sole use of the intended recipient(s).  It may 
contain information that is privileged and confidential.  Any unauthorized 
review, use, disclosure, or distribution is prohibited. If you are not the 
intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email. 


-Original Message-
From: Richard Cartun [mailto:rcar...@harthosp.org] 
Sent: Thursday, January 10, 2013 2:47 PM
To: Weems, Joyce K.; Mark Tarango; Mike Pence; Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

I'm not 100% sure, but I don't think this CPT code has a Technical component.

Richard

Richard W. Cartun, MS, PhD
Director, Histology  Immunopathology
Director, Biospecimen Collection Programs Assistant Director, Anatomic 
Pathology Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-2204 Fax


 Mike Pence mpe...@grhs.net 1/10/2013 2:27 PM 
So are you putting the charge thru twice or is the charge for the 88363 divided 
out into tech and prof.?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce K.
Sent: Thursday, January 10, 2013 1:06 PM
To: 'Mark Tarango'; Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval


And I should explain the reason I most know this.. our pathologists were denied 
because the tech charge hadn't been entered yet. So now I make sure the tech 
charge is entered before sending to the pathologists billing folks.

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org 



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's 
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It may contain information that is privileged and confidential.  Any 
unauthorized review, use, disclosure, or distribution is prohibited. If you are 
not the intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email.


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark Tarango
Sent: Thursday, January 10, 2013 2:01 PM
To: Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Help with CPT code 88363 for archived tissue retrieval

It can't be used to just pull blocks.  The slides have to be reviewed and the 
best block chosen by a pathologist.  If there is only one block then the 
pathologist needs to look at the slides and determine if there is enough tissue 
for molecular testing.  It's a professional charge.

Use it on re-accessioned cases for which molecular testing is requested and the 
best block needs to be chosen.

Yes we use it when sending out for Oncotype DX if the case was signed out over 
30 days before.

Mark

On Thu, Jan 10, 2013 at 7:36 AM, Natalie Nagy  nagy_nata...@holyokehealth.com 
wrote:

 Hi everyone,
I just have a question about CPT code 88363, first 
 can it be used for pulling blocks for Oncotype DX testing, also is 
 there a time limit on when this code can be used? Does it have to be 
 within a year, a month, etc...of when the patient account went active?

 Thanks for all the help,

 Natalie J. Nagy (HT)ASCP
 Histology Supervisor
 Holyoke Medical Center


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RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

[Weems, Joyce K.]

Same here - and we are being reimbursed. We don't charge much. Medicare new 
rate is $12.71 - but every billable test helps when that is what our 
productivity is based on! 

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's 
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intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email. 


-Original Message-
From: Victor A. Tobias [mailto:vtob...@uw.edu] 
Sent: Thursday, January 10, 2013 5:47 PM
To: 'Bell, Lynne'; 'Richard Cartun'; Weems, Joyce K.; 'Mark Tarango'; 'Mike 
Pence'; 'Natalie Nagy'
Cc: 'histonet@lists.utsouthwestern.edu'
Subject: RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

We have been billing a tech and pro fee since 2011. Whether we are getting 
reimbursed for both is another question.

Victor

Victor Tobias HT(ASCP)
Clinical Applications Analyst
Harborview Medical Center
Dept of Pathology Room NJB244
Seattle, WA 98104
vtob...@u.washington.edu
206-744-2735
206-744-8240 Fax
=
Privileged, confidential or patient identifiable information may be contained 
in this message. This information is meant only for the use of the intended 
recipients. If you are not the intended recipient, or if the message has been 
addressed to you in error, do not read, disclose, reproduce, distribute, 
disseminate or otherwise use this transmission. Instead, please notify the 
sender by reply e-mail, and then destroy all copies of the message and any 
attachments.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bell, Lynne
Sent: Thursday, January 10, 2013 11:50 AM
To: 'Richard Cartun'; Joyce K. Weems; Mark Tarango; Mike Pence; Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

I agree, Dr. Cartun.  I believe that the CPT Coding book specifically says that 
it is only a professional charge.

Lynne Bell, HT (ASCP)
Histology Team Leader
Central Vermont Medical Center
130 Fisher Road
Berlin, VT  05641
802-371-4923
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Richard Cartun
Sent: Thursday, January 10, 2013 2:47 PM
To: Joyce K. Weems; Mark Tarango; Mike Pence; Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval

I'm not 100% sure, but I don't think this CPT code has a Technical component.

Richard

Richard W. Cartun, MS, PhD
Director, Histology  Immunopathology
Director, Biospecimen Collection Programs Assistant Director, Anatomic 
Pathology Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-2204 Fax


 Mike Pence mpe...@grhs.net 1/10/2013 2:27 PM 
So are you putting the charge thru twice or is the charge for the 88363 divided 
out into tech and prof.?

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce K.
Sent: Thursday, January 10, 2013 1:06 PM
To: 'Mark Tarango'; Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help with CPT code 88363 for archived tissue retrieval


And I should explain the reason I most know this.. our pathologists were denied 
because the tech charge hadn't been entered yet. So now I make sure the tech 
charge is entered before sending to the pathologists billing folks.

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org 



www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342

This e-mail, including any attachments is the property of Saint Joseph's 
Hospital and is intended for the sole use of the intended recipient(s).
It may contain information that is privileged and confidential.  Any 
unauthorized review, use, disclosure, or distribution is prohibited. If you are 
not the intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email.


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mark Tarango
Sent: Thursday, January 10, 2013 2:01 PM
To: Natalie Nagy
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Help with CPT code 88363 for archived tissue retrieval

It can't be used to just pull blocks.  The slides have to be reviewed

Re: [Histonet] Help

[Marvin Hanna]

Hi Ly,

When you click on the Unsubscribe button, an email is sent to you to 
confirm you want to unsubscribe. It has a link to click to confirm you 
want to unsubscribe. Is the confirmation email getting lost in your junk 
folder? You won't be unsubscribed until you click the link in the 
confirmation email. The confirmation email should have a subject like, 
confirm 1a5b and a return address of 
histonet-requ...@list.utsouthwestern.edu. Hope this helps.


Best Regards,

Marvin Hanna

On 12/28/2012 2:04 PM, Ly Nguyen wrote:

I want to be remove from the list.  I've tried the unsubscribe link be low
several time but it seem like it doesn't work because I'm still receiving
daily email





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RE: [Histonet] Help

[Bartlett, Jeanine (CDC/OID/NCEZID)]

For some reason I just received 3 emails from Histonet asking me to confirm if 
I want to be unsubscribedI did not request to be unsubscribed.

Did anyone else receive emails like this?

Jeanine H. Bartlett
Centers for Disease Control and Prevention
Infectious Diseases Pathology Branch
404-639-3590
jeanine.bartl...@cdc.hhs.gov

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Marvin Hanna
Sent: Friday, December 28, 2012 2:28 PM
To: histonet@lists.utsouthwestern.edu; ln0...@gmail.com
Subject: Re: [Histonet] Help

Hi Ly,

When you click on the Unsubscribe button, an email is sent to you to confirm 
you want to unsubscribe. It has a link to click to confirm you want to 
unsubscribe. Is the confirmation email getting lost in your junk folder? You 
won't be unsubscribed until you click the link in the confirmation email. The 
confirmation email should have a subject like, confirm 1a5b and a return 
address of histonet-requ...@list.utsouthwestern.edu. Hope this helps.

Best Regards,

Marvin Hanna

On 12/28/2012 2:04 PM, Ly Nguyen wrote:
 I want to be remove from the list.  I've tried the unsubscribe link be 
 low several time but it seem like it doesn't work because I'm still 
 receiving daily email




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Re: [Histonet] Help

[Marvin Hanna]

Hi Jeanine,

Hmmm... The only way that should happen is if someone put your email 
address in and clicked unsubscribe 3 times. That's the purpose of the 
confirmation email so that no one else can unsubscribe (or subscribe) 
you. So, anyone out there trying to unsubscribe Jeanine, please stop. 
The computer server is smarter than you. And we like Jeanine. :-)


Marvin

On 12/28/2012 2:33 PM, Bartlett, Jeanine (CDC/OID/NCEZID) wrote:

For some reason I just received 3 emails from Histonet asking me to confirm if 
I want to be unsubscribedI did not request to be unsubscribed.

Did anyone else receive emails like this?

Jeanine H. Bartlett
Centers for Disease Control and Prevention
Infectious Diseases Pathology Branch
404-639-3590
jeanine.bartl...@cdc.hhs.gov

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Marvin Hanna
Sent: Friday, December 28, 2012 2:28 PM
To: histonet@lists.utsouthwestern.edu; ln0...@gmail.com
Subject: Re: [Histonet] Help

Hi Ly,

When you click on the Unsubscribe button, an email is sent to you to confirm you want to 
unsubscribe. It has a link to click to confirm you want to unsubscribe. Is the confirmation email getting 
lost in your junk folder? You won't be unsubscribed until you click the link in the confirmation email. The 
confirmation email should have a subject like, confirm 1a5b and a return address of 
histonet-requ...@list.utsouthwestern.edu. Hope this helps.

Best Regards,

Marvin Hanna

On 12/28/2012 2:04 PM, Ly Nguyen wrote:

I want to be remove from the list.  I've tried the unsubscribe link be
low several time but it seem like it doesn't work because I'm still
receiving daily email




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RE: [Histonet] Help! Liver mistakenly processed in paraffin (had tobe in OCT instead)!

[Patsy Ruegg]

Yea if you were looking for fat that will be lost with paraffin processing,
so the oil red o for fat will not work.  I would not see any point in trying
to depara and then snap freeze, but if I did want to do that since the
tissue is formalin fixed I would depara, fix some more in formalin and then
infiltrate the tissue in 30% sucrose overnight before snap freezing, if you
do not do that fix tissue will not section well frozen.

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting, LLC
40864 Arkansas Ave
Bennett, CO 80102
Phone: 303-644-4538
Fax: 720-859-4110
pru...@ihctech.net

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jennifer
MacDonald
Sent: Tuesday, November 13, 2012 10:15 AM
To: z o n k e d
Cc: histonet@lists.utsouthwestern.edu;
histonet-boun...@lists.utsouthwestern.edu
Subject: Re: [Histonet] Help! Liver mistakenly processed in paraffin (had
tobe in OCT instead)!

It depends on what you are using the oil red o for.  Lipofuscin and ceroid 
can be demonstrated with an oil red o stain after processing.
Jennifer MacDonald




From:   z o n k e d zon...@gmail.com
To: histonet@lists.utsouthwestern.edu 
histonet@lists.utsouthwestern.edu
Date:   11/13/2012 08:53 AM
Subject:[Histonet] Help! Liver mistakenly processed in paraffin 
(had to be  in OCT instead)!
Sent by:histonet-boun...@lists.utsouthwestern.edu



Hello Histonetters,

First time writer, long time reader. I'm a newbie tech in academia and I
was given a simple task which I think I pretty much screwed up.

I should have embedded half of a mouse liver in paraffin for microtome
sectioning while the other half should have been embedded in OCT for
cryosectioning (for oil red o). I made the mistake last night of placing
both liver halves into the tissue processor. The liver I intended for OCT
embedding is now hard as wax. Is there any way to deparaffinize processed
organs and may I embed them in OCT for proper cryosectioning? I imagine
that the liver would get dehydrated, I would get crappy sections, and Oil
Red O won't work.

Any suggestions are welcome.

Thank you so much,

Zoe W.


-- 
It costs nothing to say something kind. Even less to shut up altogether.

--Nathan Fillion
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Re: [Histonet] Help! Liver mistakenly processed in paraffin (had to be in OCT instead)!

[Will Chappell]

Nope, sorry. All your fat is dissolved. 

Sent from my iPhone

On Nov 13, 2012, at 8:52 AM, z o n k e d zon...@gmail.com wrote:

 Hello Histonetters,
 
 First time writer, long time reader. I'm a newbie tech in academia and I
 was given a simple task which I think I pretty much screwed up.
 
 I should have embedded half of a mouse liver in paraffin for microtome
 sectioning while the other half should have been embedded in OCT for
 cryosectioning (for oil red o). I made the mistake last night of placing
 both liver halves into the tissue processor. The liver I intended for OCT
 embedding is now hard as wax. Is there any way to deparaffinize processed
 organs and may I embed them in OCT for proper cryosectioning? I imagine
 that the liver would get dehydrated, I would get crappy sections, and Oil
 Red O won't work.
 
 Any suggestions are welcome.
 
 Thank you so much,
 
 Zoe W.
 
 
 -- 
 It costs nothing to say something kind. Even less to shut up altogether.
 
--Nathan Fillion
 ___
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Re: [Histonet] Help! Liver mistakenly processed in paraffin (had to be in OCT instead)!

[Rena Fail]

Oil Red O is a stain for fat, Alcohol dissolves fat Rehydrating  won't
help. You  can't replace the fat.
Rena Fail

On Tue, Nov 13, 2012 at 11:52 AM, z o n k e d zon...@gmail.com wrote:

 Hello Histonetters,

 First time writer, long time reader. I'm a newbie tech in academia and I
 was given a simple task which I think I pretty much screwed up.

 I should have embedded half of a mouse liver in paraffin for microtome
 sectioning while the other half should have been embedded in OCT for
 cryosectioning (for oil red o). I made the mistake last night of placing
 both liver halves into the tissue processor. The liver I intended for OCT
 embedding is now hard as wax. Is there any way to deparaffinize processed
 organs and may I embed them in OCT for proper cryosectioning? I imagine
 that the liver would get dehydrated, I would get crappy sections, and Oil
 Red O won't work.

 Any suggestions are welcome.

 Thank you so much,

 Zoe W.


 --
 It costs nothing to say something kind. Even less to shut up altogether.

 --Nathan Fillion
 ___
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 Histonet@lists.utsouthwestern.edu
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Re: [Histonet] Help! Liver mistakenly processed in paraffin (had to be in OCT instead)!

[Jennifer MacDonald]

It depends on what you are using the oil red o for.  Lipofuscin and ceroid 
can be demonstrated with an oil red o stain after processing.
Jennifer MacDonald




From:   z o n k e d zon...@gmail.com
To: histonet@lists.utsouthwestern.edu 
histonet@lists.utsouthwestern.edu
Date:   11/13/2012 08:53 AM
Subject:[Histonet] Help! Liver mistakenly processed in paraffin 
(had to be  in OCT instead)!
Sent by:histonet-boun...@lists.utsouthwestern.edu



Hello Histonetters,

First time writer, long time reader. I'm a newbie tech in academia and I
was given a simple task which I think I pretty much screwed up.

I should have embedded half of a mouse liver in paraffin for microtome
sectioning while the other half should have been embedded in OCT for
cryosectioning (for oil red o). I made the mistake last night of placing
both liver halves into the tissue processor. The liver I intended for OCT
embedding is now hard as wax. Is there any way to deparaffinize processed
organs and may I embed them in OCT for proper cryosectioning? I imagine
that the liver would get dehydrated, I would get crappy sections, and Oil
Red O won't work.

Any suggestions are welcome.

Thank you so much,

Zoe W.


-- 
It costs nothing to say something kind. Even less to shut up altogether.

--Nathan Fillion
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Re: [Histonet] Help! Liver mistakenly processed in paraffin (had to be in OCT instead)!

[Rene J Buesa]

You really screwed it up!
When you placed both pieces of liver in the processor both were subjected to 
the effect of ethanol and probably xylene and both reagents extracted the liver 
fat and no matter what you try to do now, there will be not enough fat in the 
pieces as to even try the ORO stain.
Try to get another piece. Anything you will try will not render good 
cryosections and no fat staining.
René J.



From: z o n k e d zon...@gmail.com
To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu 
Sent: Tuesday, November 13, 2012 11:52 AM
Subject: [Histonet] Help! Liver mistakenly processed in paraffin (had to be in 
OCT instead)!

Hello Histonetters,

First time writer, long time reader. I'm a newbie tech in academia and I
was given a simple task which I think I pretty much screwed up.

I should have embedded half of a mouse liver in paraffin for microtome
sectioning while the other half should have been embedded in OCT for
cryosectioning (for oil red o). I made the mistake last night of placing
both liver halves into the tissue processor. The liver I intended for OCT
embedding is now hard as wax. Is there any way to deparaffinize processed
organs and may I embed them in OCT for proper cryosectioning? I imagine
that the liver would get dehydrated, I would get crappy sections, and Oil
Red O won't work.

Any suggestions are welcome.

Thank you so much,

Zoe W.


-- 
It costs nothing to say something kind. Even less to shut up altogether.

    --Nathan Fillion
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Re: [Histonet] help with dead fungi

[Rene J Buesa]

Do you mean to try to demonstrate if the fungi you find in a sample WAS dead or 
alive before it was killed during the fixation and processing?
My take on this is the following: if a fungi dies naturally in a tissue 
(either lung or nail, or whatever) that dead fungi either is decayed and lost 
or its shape has to be modified in a way that it can be microscopically 
distinguished by the pathologist from other alive fungi.
On the other hand you could try to use DNA or RNA staining to localize the 
nuclei and decide if the structure seems to correspond to that of an alive 
fungi.
I think that if you find fungal structures in abundance in a FFPE tissue they 
should correspond to living fungi at the moment the FFPE process started.
René J.



From: Bartlett, Jeanine (CDC/OID/NCEZID) j...@cdc.gov
To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu 
Sent: Thursday, October 11, 2012 7:53 AM
Subject: [Histonet] help with dead fungi

Good morning everyone,

Does anyone have  a protocol they use to demonstrate dead fungi in FFPPE tissue?

Thanks!

Jeanine H. Bartlett, BS HT(ASCP), QIHC
Centers for Disease Control and Prevention
Infectious Diseases Pathology Branch
1600 Clifton Road, NE
MS/G-32
Atlanta, Ga 30333
404-639-3590
jeanine.bartl...@cdc.hhs.gov

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Re: [Histonet] help ! paraffin section

[Rene J Buesa]

What you describe is a typical example of poor paraffin infiltration = the 
paraffin has not infiltrated the tissue and when you prepare the final block it 
will consist of 2 different components; the tissue and the paraffin. That is 
why you end with a good paraffin section without the tissue.
Poor paraffin infiltration is always caused by an improper sequence while 
tissue processing. Either the fixation is incomplete 
OR the dehydration is incomplete and there is water in the tissue when you go 
to the clearing stage 
OR the clearing stage is incomplete and the tissue still has alcohol 
(immiscible with paraffin) when the tissue goes to the paraffin 
OR the paraffin infiltration is too short.
The problem resides in your processing protocol and there is nothing you can do 
about that at the end.
Try to check your processing protocol to eliminate the problem.
If this is happening all of the sudden while you used to have good results 
previously, then you either have changed reagents or the reagents are not in a 
good condition.
René J. 



From: Megha Kumar meg...@g.clemson.edu
To: histonet@lists.utsouthwestern.edu 
Sent: Tuesday, August 7, 2012 11:45 PM
Subject: [Histonet] help ! paraffin section

Hi All
I am trying to section adult mouse intestine and skin using paraffin
embedding. However, when i section, the tissue is torn although the rest of
the paraffin looks perfect. Please suggest why this is happening. Also,
sometimes the skin sections fall off the slides when I perform in situ
hybridization. Any ideas how to prevent this?
Please help! i am a beginner in histology and dont' know what to do!
regards
Megha


*
*
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Re: [Histonet] help ! paraffin section

[Mehmet Fatih BOZKURT]

In addition to Rene's comment,to cut coagulated tissue (skin that have
new wound crust) and calcified tissue is difficult.

On Wed, Aug 8, 2012 at 6:45 AM, Megha Kumar meg...@g.clemson.edu wrote:

 Hi All
 I am trying to section adult mouse intestine and skin using paraffin
 embedding. However, when i section, the tissue is torn although the rest of
 the paraffin looks perfect. Please suggest why this is happening. Also,
 sometimes the skin sections fall off the slides when I perform in situ
 hybridization. Any ideas how to prevent this?
 Please help! i am a beginner in histology and dont' know what to do!
 regards
 Megha


 *
 *
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-- 
Mehmet Fatih BOZKURT, DVM, PhD
Afyon Kocatepe University
Faculty of Veterinary Medicine
Department of Pathology
03030, ANS Campus
Afyonkarahisar-TURKEY
Tel: +902722281312-173/237
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Re: [Histonet] Help

[Brendal Finlay]

Nancy,


We've had similar issues with fatty tissue falling off of the slides
while performing IHC.  We use Superfrost + slides which we have found
to really hold the tissue well.  Also, I have learned through reading
round on the Histonet that air drying doesn't completely remove the
water from the middle area of a tissue section.  For this reason, we
no longer air dry at all unless it's a slide that was cut the day
before and just happened to be air dried.  


Our protocol changed to cutting the slides and draining them well,
then putting them in a 60 C oven for 15 minutes.  Then the slides
are run down to water on an automated stainer with another 15 minute
time in the oven on the stainer.  


A specific instance when the tissue falls off, was during antigen
retrieval in Trilogy in a pressure cooker.  If the pressure was
manually released, this would cause the Trilogy to boil and it would
separate the tissue from the slide.  Ourprotocol changed to 12
minutes in the pressure cooker with Trilogy, then around 8 minutes to
wait for the pressure to release on it's own.  We would then rinse
softly in distilled water to remove the Trilogy.  This also seemed
to help with the issue.

  

The combination of this has worked fairly well for us with some
exceptionally stubborn tissue still attempting to fall off of the
slides.  I would love to hear of other's experiences and how they
resolved this.  


I do wonder about the length of time in your oven.  I had spoken with
one of our Biocare reps about this when we encountered the problem and
he felt that longer than 30 minutes in the oven would damage the
specimen's IHC integrity.  


Brendal Finlay HT (ASCP)


Original message-
From: Cloughley-Gray, Nancy cloughl...@rvh.on.ca
Date: Fri, 18 May 2012 14:02:35 -0500
To:
'histonet@lists.utsouthwestern.edu'histonet@lists.utsouthwestern.edu
Subject: [Histonet] Help

 I'm a Histotechnologist working in the Regional Hospital in Barrie,
ON Canada. We are using the Ventana Ultra for our Immunohistochemistry
(IHC). Since the end of February, we have been having issues with some
tissues lifting off our positive (marked with +) charged slides. It
seems to be mostly with the fatty and/or larger sections. We now dry
our slides for one hour at room temperature (R.T.) and an additional
hour at 60 degrees C. We cut our IHC sections at 4 um. Since we have
tried 2 different types of + slides and will be trying another type of
charged slide (from Newcomer this time) I was wondering if anyone has
any other suggestions?
 I also have another question regarding a QC (quality control) issue.
We use a multi-tissue control that is applied to the top of all our
test slides for IHC. One of our paths commented that there is some
positive staining in the smooth muscle nuclei of thenormal bowel when
we are testing for Progesterone (PR). We are using a Heat Induce
Epitope Retrieval (HEIR) of 36 minutes with CC1 (Ventana's proprietary
buffer @ pH of 8.0-8.5) and a primary antibody incubation time of 16
minutes with PR clone 1E2. (Ventana instrumentation provides
pre-diluted antibodies and the user adjusts the concentration of the
antibody by adjusting the time the primary antibody is incubated with
the tissue).
 I am concerned about the implications of this staining and I have
not been able to find a reference to this kind of unusual staining
pattern. The bowel tissue that we are using as QC is from a 62 year
old female patient. I was wondering if anyone has had any experience
with this kind of staining and /or any references that I could use.
 
 Thanking you in advance,
 I look forward to your input,
 Nancy Cloughley-Gray MLT
 


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RE: [Histonet] Help

[Monfils, Paul]

I realize that such + slides come with the instruction to completely
dry slides at room temperature before placing in the drying oven.  I
have used these slides for many years, and have found this procedure to
be not only unnecessary, but sometimes problematic. I believe sections
are more likely to detach if dried at room temperature prior to oven
drying.  If the section is not lying perfectly flat against the glass -
and some types of tissue never do initially - room temperature drying
doesn't allow wrinkled areas or other problem areas to effectively
spread flat. Points that are in contact with the glass bond
electrostatically, but points that are separated from the glass, even by
a few microns, do not.  Then, when placed into the oven, such raised
areas cannot spread flat because closely adjacent areas are already
bonded to the slide, and cannot move.

After picking up sections from the waterbath, I allow them to stand
vertically and drain for no more than 5 minutes, then place them into
the drying oven at 70 degrees C. for an hour.  I very seldom have any
detachment problems with this protocol.

Paul M.


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Re: [Histonet] Help

[Rene J Buesa]

As to your issue of tissue not adhering to the slides, you could try to check 
the expiration date of your (+) slides. Perhaps it is just an issue with the 
slide.
As to controlling the concentration of an antibody by changing the incubation 
time, that is somewhat unorthodox to say the least. You modify a concentration 
with dilution, not with time. Perhaps you could modify the HIER step, or try to 
dilute the antibody.
René J.

--- On Fri, 5/18/12, Cloughley-Gray, Nancy cloughl...@rvh.on.ca wrote:


From: Cloughley-Gray, Nancy cloughl...@rvh.on.ca
Subject: [Histonet] Help
To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu
Cc: Callan, Lisa call...@rvh.on.ca
Date: Friday, May 18, 2012, 4:02 PM


I'm a Histotechnologist working in the Regional Hospital in Barrie, ON Canada. 
We are using the Ventana Ultra for our Immunohistochemistry (IHC). Since the 
end of February, we have been having issues with some tissues lifting off our 
positive (marked with +) charged slides. It seems to be mostly with the fatty 
and/or larger sections. We now dry our slides for one hour at room temperature 
(R.T.) and an additional hour at 60 degrees C. We cut our IHC sections at 4 um. 
Since we have tried 2 different types of + slides and will be trying another 
type of charged slide (from Newcomer this time) I was wondering if anyone has 
any other suggestions?
I also have another question regarding a QC (quality control) issue. We use a 
multi-tissue control that is applied to the top of all our test slides for IHC. 
One of our paths commented that there is some positive staining in the smooth 
muscle nuclei of the normal bowel when we are testing for Progesterone (PR). We 
are using a Heat Induce Epitope Retrieval (HEIR) of 36 minutes with CC1 
(Ventana's proprietary buffer @ pH of 8.0-8.5) and a primary antibody 
incubation time of 16 minutes with PR clone 1E2. (Ventana instrumentation 
provides pre-diluted antibodies and the user adjusts the concentration of the 
antibody by adjusting the time the primary antibody is incubated with the 
tissue).
I am concerned about the implications of this staining and I have not been able 
to find a reference to this kind of unusual staining pattern. The bowel tissue 
that we are using as QC is from a 62 year old female patient. I was wondering 
if anyone has had any experience with this kind of staining and /or any 
references that I could use.

Thanking you in advance,
I look forward to your input,
Nancy Cloughley-Gray MLT

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Re: [Histonet] help

[koellingr]

Hi Lydia, 

I think Tony Henwood has it exactly right in talking of DAB intensification.  
The article he sites and several others show how much more sensitive DAB 
intensification can make an ordinary iron reaction.  And you are not looking in 
bone marrow or spleen but somewhere where there is so little iron. 

  

More than once I've been burned by research colleagues who gave me tissue 
saying x should be upregulated and spend weeks looking for it until they say 
oop's, sorry-my bad. Intra-dermal injections can become sub-dermal. IP 
injections can be sub-optimal with rough handling of mice. IV tail vein 
injections can be missed. And MPTP is toxic and dangerous enough to work with 
that I'd be fumbling around and nervous myself.  If you are confident the model 
is working by some secondary marker such as tyrosine hydroxylase 
immunoreactivity then you are still are looking for minute quantities of iron. 

  

Several easily available papers on that very mouse model tell you the limited 
cell population to look for (one says-NOT in the big cells), and in a very 
specific region using coronal sections (I always used a mouse brain mold to 
section to be sure of the anatomical location) and several of the papers use 
classical iron histochemistry followed by DAB intensification and their 
procedures are in material and methods. 

  

Reaction might be working after all-just have to focus in on very limited 
reaction product.  Good luck hunting. 

  

Ray 

  

Ray Koelling 

PhenoPath Labs 

Seattle, WA 



- Original Message -




From: Lydia Gunawan lydia.guna...@unimelb.edu.au 
To: Histonet@lists.utsouthwestern.edu 
Sent: Tuesday, November 15, 2011 2:00:32 PM 
Subject: [Histonet] help 

Hi there, I am having trouble with Turnbull staining. Anybody can help me? 
As I want to detect and quantify iron in the brain for Parkinson experiment, I 
am using mice(C57Bl6)  brain that were treated with MPTP injection.  So, I have 
been trying to stain those brain using paraffin section with Turnbull blue but 
I have no luck. 
FYI, I have been using 7% of Potassium ferricyanide in 3% HCL and I incubated 
for 2 hours, 37'. I also did incubation in triton-x and H2O2 too but still not 
getting any iron on my sections. Could anyone help me to solve my problem? 
Thanks 



Lydia Gunawan 
Oxidation Biology Laboratory 
Mental Health Research Institute 
Melbourne Brain Centre 
Corner Royal Pde and Genetics Lane 
University of Melbourne, Level 4 
Parkville, Vic 3010 
email: lydia.guna...@unimelb.edu.au 


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Re: [Histonet] help

[John Kiernan]

 
 
 
 
 
 
 

The Turnbull's blue method as you describe it will not detect iron in tissues. 
 
Potassium ferricyanide will give a blue precipitate only with iron(II) 
(ferrous). In tissues, the iron is present as iron(III) (ferric) in such 
proteins as ferritin and haemosiderin. The iron of haemoglobin, though 
abundant, cannot be released by acid to react as Fe(II) or Fe(III) ions. The 
easiest way to detect Fe(III) is with potassium ferrocyanide - 0.05M in 0.2M 
HCl, for 30m - which gives a Prussian blue deposit (Perls' reaction).  If you 
want to do a Turnbull's blue method, which some say is a bit more sensitive, 
you must first reduce all the Fe(III) in the tissue to Fe(II) with dilute 
ammonium sulphide. All the iron then ends up as precipitated FeS, which will 
react with an acidified potassium ferricyanide solution to produce Turnbull's 
blue.
 
In fact, Prussian and Turnbull's blues are the same compound (see inorganic 
chemistry textbooks). A faint or invisible reaction product can be amplified 
because this pigment behaves like peroxidase, catalyzing the oxidation of DAB 
by H2O2 (see e.g. Connor et al. 1995). The sensitivity can be further increased 
with chemical tricks to change the brown oxidation product of DAB into larger 
quantities of black stuff (Moos  Mollgard 1993).
 
References. 
Connor JR et 4 al (1995) A histochemical study of iron-positive cells in the 
developing rat brain. J. Comp. Neurol. 355:111-123. 
Moos T  Mollgard K (1993) A sensitive post-DAB enhancement technique for 
demonstration of iron in the central nervous system. Histochemistry 99:471-475. 
 
John Kiernan 
Anatomy, UWO 
London, Canada 
= = =

- Original Message -
From: Lydia Gunawan lydia.guna...@unimelb.edu.au 
To: Histonet@lists.utsouthwestern.edu 
Sent: Tuesday, November 15, 2011 2:00:32 PM 
Subject: [Histonet] help 

Hi there, I am having trouble with Turnbull staining. Anybody can help me? 
As I want to detect and quantify iron in the brain for Parkinson experiment, I 
am using mice(C57Bl6)  brain that were treated with MPTP injection.  So, I have 
been trying to stain those brain using paraffin section with Turnbull blue but 
I have no luck. 
FYI, I have been using 7% of Potassium ferricyanide in 3% HCL and I incubated 
for 2 hours, 37'. I also did incubation in triton-x and H2O2 too but still not 
getting any iron on my sections. Could anyone help me to solve my problem? 
Thanks 



Lydia Gunawan 
Oxidation Biology Laboratory 
Mental Health Research Institute 
Melbourne Brain Centre 
Corner Royal Pde and Genetics Lane 
University of Melbourne, Level 4 
Parkville, Vic 3010 
email: lydia.guna...@unimelb.edu.au 


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RE: [Histonet] Help with Masson's Trichrome - not working on cryo sections

[Tony Henwood]

Jennifer,

Looking at the images, is it possible that the slides were air-dried for too 
long prior to staining?

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager  Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jennifer Fricton
Sent: Wednesday, 1 June 2011 2:28 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Help with Masson's Trichrome - not working on cryo sections

Hello,

I am completely stumped by a problem we¹ve been having with our Masson¹s 
Trichrome.  It isn¹t working properly on cryo sections.  We developed our 
protocol several years ago and it has been a standard in our lab without any 
trouble, until about four months ago when it just stopped working on frozen 
sections.  It still works fine on paraffin sections.

You can see images at this link:
http://www.med.umn.edu/lhi/prod/groups/med/@pub/@med/@dom/@lhi/documents/con
tent/med_content_340494.pdf

What seems to be happening is that the beibrich scarlet + acid fuchsin reagent 
is either not staining the muscle tissue, or is washing out of the tissue.  The 
aniline blue is still staining the connective tissue very well (it stopped 
working, too, but we solved by reducing time in water rinse following stain).  
My Masson¹s protocol is very standard ­ essentially straight out of the Armed 
Forces Institute of Pathology Manual (p. 94).  I fix my cryosections in 10% NBF 
x 30 mins. before starting the protocol.

I¹ve made sure my formalin is fresh and properly stored, with no white 
precipitate.  I¹ve tried the Bouin¹s mordant step at both 56C and room temp 
overnight.  I¹ve reduced my distilled water rinses following the biebrich 
scarlet and aniline blue steps to just a quick dip, thinking maybe I was 
de-staining too much before setting the stains with acetic acid.  I¹ve re-made 
all solutions, checked all components and pH levels.  I haven¹t done anything 
different with my frozen tissues, and I¹ve tried several different freshly cut 
tissues.

Has anyone seen this before and have any tips on what is going wrong?  Your 
help is greatly appreciated!

--
Jennifer L. Fricton, B.S.
Scientist, Lab Manager
Lillehei Heart Institute Histology  Microscopy Core Facility University of 
Minnesota School of Medicine, Division of Cardiology
312 Church Street SE
4-290 Nils Hasselmo Hall
Minneapolis, MN  55455
Lab: 612-626-3090

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RE: [Histonet] [Help] oil red O stain in fattty change liver

[Setlak, Lisa]

We do this stain on frozen sections in our lab with no formalin fixation. We 
stain in oil red o from American Mastertech for 30 minutes, rinse in water, 
counterstain in hematoxylin, coverslip with aqueous mounting media.
Lisa

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of kyoung LEE
Sent: Thursday, April 14, 2011 9:07 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] [Help] oil red O stain in fattty change liver

Dear sirs.

I don't know whether I send this question to you.

For several weeks, I'm setting oil red O stain in fattty change liver (mouse).

Could you review my protocol?

1. Murine tissue,livers, is fixed with 10% NBF(10% neutral-buffered formalin).
2. Make frozen block 
3. Blocks were cut 5um in crystat and dried for 1~2H at RT. 
4. store at -20 
5. let it dry for 30mins at RT
6. immerse it in cold 10% NBF for 10mins
7. staining

After staining, my section were fallen off and seperated.

I search many protocols. Some suggest additional fixation is necessary as soon 
as cutting.
Others suggest prefixed tissue is not necessary additional fixation.

Plz send me your comments.
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Re: [Histonet] [Help] oil red O stain in fatty change liver

[Jennifer MacDonald]

We use pre-fixed tissue for oil red o.  We rinse the section briefly to 
remove the formalin from the outside of the tissue.  We prepare our tissue 
in OCT or the equivalent.  We cut the frozen sections and mount them on 
charged slides.  We let dry the dry a minimum of 10 minute, rinse them in 
water to remove the OCT and then proceed with the stain.




kyoung LEE eas...@yahoo.co.kr 
Sent by: histonet-boun...@lists.utsouthwestern.edu
04/14/2011 08:04 AM

To
histonet@lists.utsouthwestern.edu
cc

Subject
[Histonet] [Help] oil red O stain in fattty change liver






Dear sirs.

I don't know whether I send this question to you.

For several weeks, I'm setting oil red O stain in fattty change liver 
(mouse).

Could you review my protocol?

1. Murine tissue,livers, is fixed with 10% NBF(10% neutral-buffered 
formalin).
2. Make frozen block 
3. Blocks were cut 5um in crystat and dried for 1~2H at RT. 
4. store at -20 
5. let it dry for 30mins at RT
6. immerse it in cold 10% NBF for 10mins
7. staining

After staining, my section were fallen off and seperated.

I search many protocols. Some suggest additional fixation is necessary as 
soon 
as cutting.
Others suggest prefixed tissue is not necessary additional fixation.

Plz send me your comments.
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RE: [Histonet] HELP!!!!! H. Pylori Immunos

[McMahon, Loralee A]

We usually run them on our automated stainers but I have been known to do them 
by hand in a pinch

Loralee McMahon, HTL (ASCP)
Immunohistochemistry Supervisor
Strong Memorial Hospital
Department of Surgical Pathology
(585) 275-7210

From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Heather Cooper 
[hctrup...@att.net]
Sent: Tuesday, March 01, 2011 12:50 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] HELP! H. Pylori Immunos

Does anyone do H. Pylori Immunos by hand?
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Re: [Histonet] HELP!!!!! H. Pylori Immunos

[Jay Lundgren]

Our problem, in my opinion is a control issue and not a staining one..our
pathologist wants to see the bugs in certain areas-glands(spelling
appologies!) its called, the circular groups of cells, in the middle of
these areas and not just occasional bugs in other areasEverywhere else
I've worked the pathologist would use a Diff-Quick, Cresyl Violet or Geimsa
in addition to the HE,   I'm aware of other newer special stains for
H.Pylori that are out there, but this pathologist is firm on the IHC method
of choice.
I use NovaCastra polyclonal, incubation @23 min. HIER with low PH for 30
minutes.with  recommended detection kit..


 Heather,

  I hope you don't mind that I copied your email so that everyone on
this forum can learn from your question.  I always use the Reply to All
button so everyone can see my response.

 I was willing to bet before I read your query that we were dealing with
a control problem, not a staining problem.  I have seen some miserable
commercial *H. pylori* controls lately.  Some areas of the country are awash
in HP, some labs can't get a good positive control block of their own.  You
must first obtain a KNOWN POSITIVE CONTROL block. Stain slides from that
block with whichever chemical special stain is acceptable to your
Pathologist, I like a modified Warthin-Starry.  Sit down with your
Pathologist and evaluate the control slides.  Is it acceptable to the
Pathologist? If so, stain more slides from the positive control block, this
time immuno stain and chemical special stain in parallel.  Evaluate the
slides with the Pathologist.  Is there correlation between the chemical and
special stain?  If so, proceed, running your first 20 cases in parallel
(immuno and chemical special stains) and documenting (with the Pathologist's
signature) the correlation of your first 20 cases.

 If you are able to demonstrate HP with a chemical special stain, but
not with your immuno procedure, call your immuno vendor.

 If you are unable to demonstrate HP with a chemical special stain, it's
your control.  Or it could be your stain, but a Giemsa is pretty hard to
mess up.

Good Luck,



Jay A. Lundgren M.S., HTL (ASCP)
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RE: [Histonet] HELP!!!!! H. Pylori Immunos

[McMahon, Loralee A]

We also cut our controls in sequence.  Stain the first of the sequence and the 
last.  To make sure that you have not cut through the area.

Loralee McMahon, HTL (ASCP)
Immunohistochemistry Supervisor
Strong Memorial Hospital
Department of Surgical Pathology
(585) 275-7210


From: Jay Lundgren [jaylundg...@gmail.com]
Sent: Tuesday, March 01, 2011 3:10 PM
To: McMahon, Loralee A
Cc: Heather Cooper; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] HELP! H. Pylori Immunos






Our problem, in my opinion is a control issue and not a staining one..our 
pathologist wants to see the bugs in certain areas-glands(spelling appologies!) 
its called, the circular groups of cells, in the middle of these areas and not 
just occasional bugs in other areasEverywhere else I've worked the 
pathologist would use a Diff-Quick, Cresyl Violet or Geimsa in addition to the 
HE,   I'm aware of other newer special stains for H.Pylori that are out there, 
but this pathologist is firm on the IHC method of choice.
I use NovaCastra polyclonal, incubation @23 min. HIER with low PH for 30 
minutes.with  recommended detection kit..


 Heather,

  I hope you don't mind that I copied your email so that everyone on this 
forum can learn from your question.  I always use the Reply to All button so 
everyone can see my response.

 I was willing to bet before I read your query that we were dealing with a 
control problem, not a staining problem.  I have seen some miserable commercial 
H. pylori controls lately.  Some areas of the country are awash in HP, some 
labs can't get a good positive control block of their own.  You must first 
obtain a KNOWN POSITIVE CONTROL block. Stain slides from that block with 
whichever chemical special stain is acceptable to your Pathologist, I like a 
modified Warthin-Starry.  Sit down with your Pathologist and evaluate the 
control slides.  Is it acceptable to the Pathologist? If so, stain more slides 
from the positive control block, this time immuno stain and chemical special 
stain in parallel.  Evaluate the slides with the Pathologist.  Is there 
correlation between the chemical and special stain?  If so, proceed, running 
your first 20 cases in parallel (immuno and chemical special stains) and 
documenting (with the Pathologist's signature) the correlation of your first 20 
cases.

 If you are able to demonstrate HP with a chemical special stain, but not 
with your immuno procedure, call your immuno vendor.

 If you are unable to demonstrate HP with a chemical special stain, it's 
your control.  Or it could be your stain, but a Giemsa is pretty hard to mess 
up.
  Good 
Luck,



 Jay A. Lundgren M.S., HTL (ASCP)
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RE: [Histonet] Help with OCT problem

[Liz Chlipala]

Donna

I have seen soft or rather sticky OCT samples in the past.  If you
freeze in isopentane and don't let the isopentane evaporate off the
samples prior to wrapping the sample in foil, that excess isopentane
changes the OCT, it makes it sticky.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
Reynolds,Donna M
Sent: Thursday, February 10, 2011 12:39 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: [Histonet] Help with OCT problem

Has anyone ever experienced OCT blocks that cut like they are soft. I
have turned the cyrostat down 10 degrees lower than usual and they are
still soft. Really weird part is that I have 20 blocks in this group and
some are soft and others are just fine. Tried changing angle, new blade
nothing helps. When I finally manage to get a decent section and pick it
up on the slide, the OCT around the tissue want even pick up smooth it
wrinkles really bad.
I tried just a block of OCT that I froze on a chuck and it cut great.  I
am clueless as to what is going on.
Samples were frozen by someone else and brought to me to be cut.
Donna Reynolds Core IHC Lab
M.D. Anderson Cancer Center, Houston TX
Research lab
713-192-8106.


Donna Reynolds Core IHC Lab
Dept. Cancer Biology, SRB 1.660
713-792-8106

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Re: [Histonet] Help with OCT problem

[Patrick Laurie]

Donna,
I would also check to make sure that the tissue wasn't held in an
something with a lot of alcohol, like RNA later.  I found out first
hand that it doesn't work well.  Good luck

On Thu, Feb 10, 2011 at 11:38 AM, Reynolds,Donna M
dreyn...@mdanderson.org wrote:
 Has anyone ever experienced OCT blocks that cut like they are soft. I have 
 turned the cyrostat down 10 degrees lower than usual and they are still soft. 
 Really weird part is that I have 20 blocks in this group and some are soft 
 and others are just fine. Tried changing angle, new blade nothing helps. When 
 I finally manage to get a decent section and pick it up on the slide, the OCT 
 around the tissue want even pick up smooth it wrinkles really bad.
 I tried just a block of OCT that I froze on a chuck and it cut great.  I am 
 clueless as to what is going on.
 Samples were frozen by someone else and brought to me to be cut.
 Donna Reynolds Core IHC Lab
 M.D. Anderson Cancer Center, Houston TX
 Research lab
 713-192-8106.


 Donna Reynolds Core IHC Lab
 Dept. Cancer Biology, SRB 1.660
 713-792-8106

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-- 
Patrick Laurie HT(ASCP)QIHC
CellNetix Pathology  Laboratories
1124 Columbia Street, Suite 200
Seattle, WA 98104
plau...@cellnetix.com

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Re: [Histonet] Help with OCT problem

[Merced M Leiker]

I agree with previous responders to this query, as I've gotten alcohol on 
my blocks and since alcohol doesn't freeze real well it makes a sticky mess 
with the OCT. The affected OCT can be scraped off with a razor blade. 
Hopefully the tissue itself did not come in contact with alcohol prior to 
embedding.


Regards,
Merced

--On Thursday, February 10, 2011 12:51 PM -0800 Patrick Laurie 
foreig...@gmail.com wrote:



Donna,
I would also check to make sure that the tissue wasn't held in an
something with a lot of alcohol, like RNA later.  I found out first
hand that it doesn't work well.  Good luck

On Thu, Feb 10, 2011 at 11:38 AM, Reynolds,Donna M
dreyn...@mdanderson.org wrote:

Has anyone ever experienced OCT blocks that cut like they are soft. I
have turned the cyrostat down 10 degrees lower than usual and they are
still soft. Really weird part is that I have 20 blocks in this group and
some are soft and others are just fine. Tried changing angle, new blade
nothing helps. When I finally manage to get a decent section and pick it
up on the slide, the OCT around the tissue want even pick up smooth it
wrinkles really bad. I tried just a block of OCT that I froze on a chuck
and it cut great.  I am clueless as to what is going on. Samples were
frozen by someone else and brought to me to be cut. Donna Reynolds Core
IHC Lab
M.D. Anderson Cancer Center, Houston TX
Research lab
713-192-8106.


Donna Reynolds Core IHC Lab
Dept. Cancer Biology, SRB 1.660
713-792-8106

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CellNetix Pathology  Laboratories
1124 Columbia Street, Suite 200
Seattle, WA 98104
plau...@cellnetix.com

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Merced M Leiker
Research Technician III
Cardiovascular Medicine
348 Biomedical Research Building
State University of New York at Buffalo
3435 Main St, Buffalo, NY 14214  USA
lei...@buffalo.edu
716-829-6118 (Ph)
716-829-2665 (Fx)

No trees were harmed in the sending of this email.
However, many electrons were severely inconvenienced.


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RE: [Histonet] Help with OCT problem

[Ingles Claire]

Same with ETOH. We wipe our surfaces down with it to clean them so no cross 
contamination as we freeze our skin here on the metal side areas of our 
cryostats. If the alcohol hasn't been wiped dry or evaporated we have that 
problem too.
Claire



From: histonet-boun...@lists.utsouthwestern.edu on behalf of Liz Chlipala
Sent: Thu 2/10/2011 1:49 PM
To: Reynolds,Donna M; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help with OCT problem



Donna

I have seen soft or rather sticky OCT samples in the past.  If you
freeze in isopentane and don't let the isopentane evaporate off the
samples prior to wrapping the sample in foil, that excess isopentane
changes the OCT, it makes it sticky.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949
fax (303) 682-9060
www.premierlab.com





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Re: [Histonet] Help

[Joseph Saby]

Kathy-

What do these cracks look like?  Are they arranged in a parallel manner?  Or do 
they have the appearance of the cracks seen in dry mud?

Parallel aligned cracks are often found in overprocessed small biospies.  These 
small samples become hard and brittle.  The impact of the tissue on the 
microtome blade during aggressive facing force these cracks deep into the 
tissue.  With care, patience and some luck you may be able to get beyond these 
cracks by soaking the blocks, facing with thin sections on a repeated basis.  
What will determine whether you are successful will be whether there is enough 
depth to your biopsy to allow you to get beyond the facing artifact.

If the cracks resemble dry earth, then we are looking at a fixation/processing 
issue.  These cracks do not appear in the tissue until the xylenes after slide 
staining.  If the tissues are not well fixed, then processing reagents will not 
be able to fully penetrate the tissues.  If possible, I would suggest you 
perform retrims on your tissue and process them normally.  They should have had 
enough time to fix when you do your retrims, and they should be fine.  In a 
hospital setting, this may not be possible.  You can deparaffinze and rehydrate 
your tissue samples by running them through your processor's cleaning program.  
Place them back in fixative for a while, then reprocess them.  


I have also seen this artifcat in tissue when (due to a processor malfunction) 
tissue samples were exposed to high heat during processing.  Look for blood 
cells being laked in the larger blood vessels (blood cells look like a 
homogenous mass rather than being able to cell boundaries).  Sometimes there 
will be small round black precitate granules over the affected tissue areas.  
You will need to determine the best course depending on the extent of the 
damage.  Extensive damage will probaly require retrims. 

I hope this helps.  Please get back with me if yoiu have any further questions.

Joe Saby, BA HT
37 years in histotechnology



From: Kathy Nelson kathyenel...@hotmail.com
To: Histonet histonet@lists.utsouthwestern.edu
Sent: Sun, December 19, 2010 1:25:54 AM
Subject: [Histonet] Help


Solutions for cracks in tissue microscopically esp. in tumors and BCC specimens.
Thanks 
kathyenel...@hotmail.com                         
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Re: [Histonet] Help! Curled cryosections

[Teresa Iglesias]

Thanks, Montina
The sections are 40um and I had no trouble with all my other brains
(sectioned myself). I had help with these last two and they are all curled.
I think they just got sectioned too fast or something. I'll try the shaker
and see if it helps.
Thanks!

-Teresa


On Wed, Sep 15, 2010 at 8:47 PM, Montina Van Meter 
montina.vanme...@pbrc.edu wrote:

 Teresa,
 I would put the sections through several 5-10 min. washes on a shaker
 table. It is very important to make sure you have all of the cryoprotectant
 rinsed out of the tissue or it will inhibit IHC staining.  How thick are the
 sections?  Were they cut on a cryostat or freezing microtome?  I routinely
 cut rat brain at 40um and don't have any curling issues.  That sometimes
 occurs when the knife has come through the section of brain too rapidly.  A
 slow and steady motion is needed when cutting frozens.

 Good luck!
 Tina Van Meter


 Sent from my iPhone

 On Sep 15, 2010, at 9:37 PM, Teresa Iglesias tligles...@ucdavis.edu
 wrote:





-- 
__
Teresa Iglesias
Graduate Group in Animal Behavior
Department of Evolution and Ecology

University of California-Davis
One Shields Avenue
2320 Storer Hall
Davis, CA 95616
Office: 530-754-7837
Fax: 530-752-1449
tligles...@ucdavis.edu
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RE: [Histonet] Help! In need of positive Gram Control

[Susan.Walzer]

Take some fresh tissue (we use umbilical cord)  to micro and let them incubate 
in for a few days in gram + and/or negative. Then fix it and you will have good 
controls.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of connie grubaugh
Sent: Tuesday, June 22, 2010 9:06 PM
To: cg...@marylandgeneral.org; jcbrit...@cheshire-med.com; dianar...@aol.com; 
histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help! In need of positive Gram Control


Tried the slim jim and all of my doctors did not like it.  Don't waste your 
time.



Connie G.



 

 Date: Tue, 22 Jun 2010 10:16:14 -0400
 From: cg...@marylandgeneral.org
 To: jcbrit...@cheshire-med.com; dianar...@aol.com; 
 histonet@lists.utsouthwestern.edu
 Subject: RE: [Histonet] Help! In need of positive Gram Control
 CC: 
 
 You have got to be kidding!! That's hysterical. So process a slim jim
 and you have 
 Gram - and + controls. If you're serious I'm trying it. 
 
 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Josie
 Britton
 Sent: Tuesday, June 22, 2010 6:10 AM
 To: dianar...@aol.com; histonet@lists.utsouthwestern.edu
 Subject: RE: [Histonet] Help! In need of positive Gram Control
 
 
 
 
 
 Have you tried a Slim Jim? They have gram positive and negative rods in
 
 them. Regardless, I still enjoy eating them once and a while! 
 
 
 
 
 
 
 
 Josie Britton Ht
 
 
 
 Cheshire Medical Center
 
 
 
 Keene, NH 03431
 
 
 
 
 
 
 
 
 
 
 
 -Original Message-
 
 From: histonet-boun...@lists.utsouthwestern.edu
 
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
 
 dianar...@aol.com
 
 Sent: Monday, June 21, 2010 7:43 PM
 
 To: histonet@lists.utsouthwestern.edu
 
 Subject: [Histonet] Help! In need of positive Gram Control
 
 
 
 
 
 
 
 Help! We are in need of positive Gram Control Blocks if anyone has any 
 
 
 
 extra they are willing to part with. I have lots of Fungus,
 
 Pneumocystis and 
 
 
 
 HPV tissue blocks to trade.
 
 
 
 
 
 
 
 Diana Ripley
 
 
 
 John Muir Histology
 
 
 
 Concord Campus
 
 
 
 2540 East Street
 
 
 
 Concord, CA 94520
 
 
 
 ___
 
 
 
 Histonet mailing list
 
 
 
 Histonet@lists.utsouthwestern.edu
 
 
 
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 
 
 
 
 CONFIDENTIALITY NOTICE: This electronic message, including any
 attachments, is for the sole use of the intended recipients and may
 contain confidential and privileged information. Any unauthorized
 review, use, disclosure or distribution is prohibited. If you are not
 the intended recipient, please contact the sender by electronic mail and
 destroy all copies of the original message.
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RE: [Histonet] Help! In need of positive Gram Control

[Josie Britton]

It's true!  Who knew that Gram + and- riddles meat sticks could be so
yummy and versatile too! 

 

Josie

 

-Original Message-
From: Gill, Caula A. [mailto:cg...@marylandgeneral.org] 
Sent: Tuesday, June 22, 2010 10:16 AM
To: Josie Britton; dianar...@aol.com; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help! In need of positive Gram Control

 

You have got to be kidding!! That's hysterical. So process a slim jim

and you have 

Gram - and + controls. If you're serious I'm trying it. 

 

-Original Message-

From: histonet-boun...@lists.utsouthwestern.edu

[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Josie

Britton

Sent: Tuesday, June 22, 2010 6:10 AM

To: dianar...@aol.com; histonet@lists.utsouthwestern.edu

Subject: RE: [Histonet] Help! In need of positive Gram Control

 

 

 

 

 

Have you tried a Slim Jim?  They have gram positive and negative rods in

 

them.  Regardless, I still enjoy eating them once and a while! 

 

 

 

 

 

 

 

Josie Britton Ht

 

 

 

Cheshire Medical Center

 

 

 

Keene, NH 03431

 

 

 

 

 

 

 

 

 

 

 

-Original Message-

 

From: histonet-boun...@lists.utsouthwestern.edu

 

[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of

 

dianar...@aol.com

 

Sent: Monday, June 21, 2010 7:43 PM

 

To: histonet@lists.utsouthwestern.edu

 

Subject: [Histonet] Help! In need of positive Gram Control

 

 

 

 

 

 

 

Help! We are in need of positive Gram Control Blocks if anyone has any  

 

 

 

extra they are willing to part with.  I have lots of Fungus,

 

Pneumocystis  and 

 

 

 

HPV tissue blocks to trade.

 

 

 

 

 

 

 

Diana Ripley

 

 

 

John Muir Histology

 

 

 

Concord Campus

 

 

 

2540 East Street

 

 

 

Concord, CA 94520

 

 

 

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Histonet mailing list

 

 

 

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http://lists.utsouthwestern.edu/mailman/listinfo/histonet

 

 

 

 

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RE: [Histonet] Help Resurrecting Dried Fixed Tissue

[Tony Henwood]

We have success with Sandison Method, see below:


Sandison Method

Solutions:
95% ethanol 300ml
38% Formalin10ml
Sodium carbonate10g
Water   690ml

Method:

Leave tissues in solution overnight or until they become soft. Replace
one third of fluid with 95% ethanol, repeat until tissue becomes firm.


Reference:

The histological examination of Mummified Tissue Stain Techn
30:277-283, 1955

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager  Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 




-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Parsons,
Catherine
Sent: Thursday, 24 June 2010 1:16 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Help Resurrecting Dried Fixed Tissue


I am grasping at straws and hoping for ideas to salvage some formalin
fixed tissue specimens.  I work in a research lab, and am quite new to
histology and this particular lab.  A former student left formalin fixed
tissue specimens that were stored in 70% ethanol.  Unfortunately, the
seal failed on a few of the vials, the ethanol evaporated, and the
tissue is a dried out disaster at the bottom of the vial.

Has anyone has any experience / luck rehydrating tissues?  Really, any
information that I could possibly pull out of these specimens would be
welcome.

Cathy Parsons


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RE: [Histonet] Help! In need of positive Gram Control

[Gill, Caula A.]

You have got to be kidding!! That's hysterical. So process a slim jim
and you have 
Gram - and + controls. If you're serious I'm trying it. 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Josie
Britton
Sent: Tuesday, June 22, 2010 6:10 AM
To: dianar...@aol.com; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help! In need of positive Gram Control

 



Have you tried a Slim Jim?  They have gram positive and negative rods in

them.  Regardless, I still enjoy eating them once and a while! 



 



Josie Britton Ht



Cheshire Medical Center



Keene, NH 03431



 



 



-Original Message-

From: histonet-boun...@lists.utsouthwestern.edu

[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of

dianar...@aol.com

Sent: Monday, June 21, 2010 7:43 PM

To: histonet@lists.utsouthwestern.edu

Subject: [Histonet] Help! In need of positive Gram Control



 



Help! We are in need of positive Gram Control Blocks if anyone has any  



extra they are willing to part with.  I have lots of Fungus,

Pneumocystis  and 



HPV tissue blocks to trade.



 



Diana Ripley



John Muir Histology



Concord Campus



2540 East Street



Concord, CA 94520



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Re: [Histonet] Help! In need of positive Gram Control

[Victor Tobias]

What a waste of a good Slim Jim.

Victor

Victor Tobias
Clinical Applications Analyst
University of Washington Medical Center
Dept of Pathology Room BB220
1959 NE Pacific
Seattle, WA 98195
vic...@pathology.washington.edu
206-598-2792
206-598-7659 Fax
=
Privileged, confidential or patient identifiable information may be
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transmission. Instead, please notify the sender by reply e-mail, and
then destroy all copies of the message and any attachments.


On 6/22/2010 7:16 AM, Gill, Caula A. wrote:

You have got to be kidding!! That's hysterical. So process a slim jim
and you have
Gram - and + controls. If you're serious I'm trying it.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Josie
Britton
Sent: Tuesday, June 22, 2010 6:10 AM
To: dianar...@aol.com; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Help! In need of positive Gram Control





Have you tried a Slim Jim?  They have gram positive and negative rods in

them.  Regardless, I still enjoy eating them once and a while!







Josie Britton Ht



Cheshire Medical Center



Keene, NH 03431











-Original Message-

From: histonet-boun...@lists.utsouthwestern.edu

[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of

dianar...@aol.com

Sent: Monday, June 21, 2010 7:43 PM

To: histonet@lists.utsouthwestern.edu

Subject: [Histonet] Help! In need of positive Gram Control







Help! We are in need of positive Gram Control Blocks if anyone has any



extra they are willing to part with.  I have lots of Fungus,

Pneumocystis  and



HPV tissue blocks to trade.







Diana Ripley



John Muir Histology



Concord Campus



2540 East Street



Concord, CA 94520



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Re: [Histonet] Help! In need of positive Gram Control

[Jackie M O'Connor]

A good ol' hot appendix works great.   Not as good as a Slim Jim, tho.
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RE: [Histonet] Help! In need of positive Gram Control

[connie grubaugh]

Tried the slim jim and all of my doctors did not like it.  Don't waste your 
time.



Connie G.



 

 Date: Tue, 22 Jun 2010 10:16:14 -0400
 From: cg...@marylandgeneral.org
 To: jcbrit...@cheshire-med.com; dianar...@aol.com; 
 histonet@lists.utsouthwestern.edu
 Subject: RE: [Histonet] Help! In need of positive Gram Control
 CC: 
 
 You have got to be kidding!! That's hysterical. So process a slim jim
 and you have 
 Gram - and + controls. If you're serious I'm trying it. 
 
 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Josie
 Britton
 Sent: Tuesday, June 22, 2010 6:10 AM
 To: dianar...@aol.com; histonet@lists.utsouthwestern.edu
 Subject: RE: [Histonet] Help! In need of positive Gram Control
 
 
 
 
 
 Have you tried a Slim Jim? They have gram positive and negative rods in
 
 them. Regardless, I still enjoy eating them once and a while! 
 
 
 
 
 
 
 
 Josie Britton Ht
 
 
 
 Cheshire Medical Center
 
 
 
 Keene, NH 03431
 
 
 
 
 
 
 
 
 
 
 
 -Original Message-
 
 From: histonet-boun...@lists.utsouthwestern.edu
 
 [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
 
 dianar...@aol.com
 
 Sent: Monday, June 21, 2010 7:43 PM
 
 To: histonet@lists.utsouthwestern.edu
 
 Subject: [Histonet] Help! In need of positive Gram Control
 
 
 
 
 
 
 
 Help! We are in need of positive Gram Control Blocks if anyone has any 
 
 
 
 extra they are willing to part with. I have lots of Fungus,
 
 Pneumocystis and 
 
 
 
 HPV tissue blocks to trade.
 
 
 
 
 
 
 
 Diana Ripley
 
 
 
 John Muir Histology
 
 
 
 Concord Campus
 
 
 
 2540 East Street
 
 
 
 Concord, CA 94520
 
 
 
 ___
 
 
 
 Histonet mailing list
 
 
 
 Histonet@lists.utsouthwestern.edu
 
 
 
 http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 
 
 
 
 CONFIDENTIALITY NOTICE: This electronic message, including any
 attachments, is for the sole use of the intended recipients and may
 contain confidential and privileged information. Any unauthorized
 review, use, disclosure or distribution is prohibited. If you are not
 the intended recipient, please contact the sender by electronic mail and
 destroy all copies of the original message.
 ___
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RE: [Histonet] Help! In need of positive Gram Control

[Shirley A. Powell]

Go to your local drive in mart and buy you a Slim Jim, cut it in thin slices 
and you will have all the Gram controls you can use.  You will never eat 
another Slim Jim.  If you don't know what they are, they are sort of like a 
summer sausage, only thinner and guys love them.  They are found usually right 
beside the check out register.  

Shirley


From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of dianar...@aol.com 
[dianar...@aol.com]
Sent: Monday, June 21, 2010 7:43 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Help! In need of positive Gram Control

Help! We are in need of positive Gram Control Blocks if anyone has any
extra they are willing to part with.  I have lots of Fungus, Pneumocystis  and
HPV tissue blocks to trade.

Diana Ripley
John Muir Histology
Concord Campus
2540 East Street
Concord, CA 94520
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RE: [Histonet] help

[Weems, Joyce]

I think there is none... But thanks for the funny!! j 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg
Sent: Tuesday, March 23, 2010 12:00
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] help

After you stop laughing seriously I need some help here, apparently I got my 
fingers in some silver nitrate yesterday and touched my face under my nose over 
my lip and now I have black spots that won't come off.  I have done this on my 
hands before but never on my face.  So far I have tried soaking a cloth in 
hydrogen peroxide hoping to bleach it with no luck, I even tried putting some 
gold chloride on it to see if I could tone it down, to no avail.  Any ideas?  
Make up only goes so far and lasts so long.

 

Regards,

 

Patsy

 

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net

 


This email is confidential and intended solely for the use of the Person(s) 
('the intended recipient') to whom it was addressed. Any views or opinions 
presented are solely those of the author. It may contain information that is 
privileged  confidential within the meaning of applicable law. Accordingly any 
dissemination, distribution, copying, or other use of this message, or any of 
its contents, by any person other than the intended recipient may constitute a 
breach of civil or criminal law and is strictly prohibited. If you are NOT the 
intended recipient please contact the sender and dispose of this e-mail as soon 
as possible.

 

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Re: [Histonet] help

[Drew Meyer]

So sorry... the only thing I know to fix it is time... about a week or
two... :)

Drew

On Tue, Mar 23, 2010 at 11:59, Patsy Ruegg pru...@ihctech.net wrote:

 After you stop laughing seriously I need some help here, apparently I got
 my
 fingers in some silver nitrate yesterday and touched my face under my nose
 over my lip and now I have black spots that won't come off.  I have done
 this on my hands before but never on my face.  So far I have tried soaking
 a
 cloth in hydrogen peroxide hoping to bleach it with no luck, I even tried
 putting some gold chloride on it to see if I could tone it down, to no
 avail.  Any ideas?  Make up only goes so far and lasts so long.



 Regards,



 Patsy



 Patsy Ruegg, HT(ASCP)QIHC
 IHCtech, LLC
 Fitzsimmons BioScience Park
 12635 Montview Blvd. Suite 215
 Aurora, CO 80010
 P-720-859-4060
 F-720-859-4110
 wk email pru...@ihctech.net
 web site www.ihctech.net




 This email is confidential and intended solely for the use of the Person(s)
 ('the intended recipient') to whom it was addressed. Any views or opinions
 presented are solely those of the author. It may contain information that
 is
 privileged  confidential within the meaning of applicable law. Accordingly
 any dissemination, distribution, copying, or other use of this message, or
 any of its contents, by any person other than the intended recipient may
 constitute a breach of civil or criminal law and is strictly prohibited. If
 you are NOT the intended recipient please contact the sender and dispose of
 this e-mail as soon as possible.



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Re: [Histonet] help

[Emily Sours]

You could use a sharpie to fill in a killer moustache.
I say go with handlebars and not with Hitler-style.

Emily

Shall we always be content with the ancient tinned salad of the subsidized
novel? Or the tired ice-cream of poems which cry themselves to sleep in the
refrigerators of the mind?
-Lawrence Durrell, Clea


On Tue, Mar 23, 2010 at 11:59 AM, Patsy Ruegg pru...@ihctech.net wrote:

 After you stop laughing seriously I need some help here, apparently I got
 my
 fingers in some silver nitrate yesterday and touched my face under my nose
 over my lip and now I have black spots that won't come off.  I have done
 this on my hands before but never on my face.  So far I have tried soaking
 a
 cloth in hydrogen peroxide hoping to bleach it with no luck, I even tried
 putting some gold chloride on it to see if I could tone it down, to no
 avail.  Any ideas?  Make up only goes so far and lasts so long.



 Regards,



 Patsy


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Re: [Histonet] help

[Paula Pierce]

You could go get a set of Groucho glasses with the attached mustache and tell 
everyone you are just gearing up for April Fool's Day!

or treat yourself to a facial and get a chemical peel.





From: Emily Sours talulahg...@gmail.com
To: Patsy Ruegg pru...@ihctech.net; histonet@lists.utsouthwestern.edu
Sent: Tue, March 23, 2010 11:09:35 AM
Subject: Re: [Histonet] help

You could use a sharpie to fill in a killer moustache.
I say go with handlebars and not with Hitler-style.

Emily

Shall we always be content with the ancient tinned salad of the subsidized
novel? Or the tired ice-cream of poems which cry themselves to sleep in the
refrigerators of the mind?
-Lawrence Durrell, Clea


On Tue, Mar 23, 2010 at 11:59 AM, Patsy Ruegg pru...@ihctech.net wrote:

 After you stop laughing seriously I need some help here, apparently I got
 my
 fingers in some silver nitrate yesterday and touched my face under my nose
 over my lip and now I have black spots that won't come off.  I have done
 this on my hands before but never on my face.  So far I have tried soaking
 a
 cloth in hydrogen peroxide hoping to bleach it with no luck, I even tried
 putting some gold chloride on it to see if I could tone it down, to no
 avail.  Any ideas?  Make up only goes so far and lasts so long.



 Regards,



 Patsy


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RE: [Histonet] help

[Edwards, Richard E.]

Suggest you Google  Henna tattoos and  then  make  a  feature  of  it with a 
few tasteful scrolls and patterns.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg
Sent: 23 March 2010 16:00
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] help

After you stop laughing seriously I need some help here, apparently I got my
fingers in some silver nitrate yesterday and touched my face under my nose
over my lip and now I have black spots that won't come off.  I have done
this on my hands before but never on my face.  So far I have tried soaking a
cloth in hydrogen peroxide hoping to bleach it with no luck, I even tried
putting some gold chloride on it to see if I could tone it down, to no
avail.  Any ideas?  Make up only goes so far and lasts so long.

 

Regards,

 

Patsy

 

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net

 


This email is confidential and intended solely for the use of the Person(s)
('the intended recipient') to whom it was addressed. Any views or opinions
presented are solely those of the author. It may contain information that is
privileged  confidential within the meaning of applicable law. Accordingly
any dissemination, distribution, copying, or other use of this message, or
any of its contents, by any person other than the intended recipient may
constitute a breach of civil or criminal law and is strictly prohibited. If
you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.

 

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RE: [Histonet] help

[Nails, Felton]

Basically time 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg
Sent: Tuesday, March 23, 2010 11:00 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] help

After you stop laughing seriously I need some help here, apparently I got my 
fingers in some silver nitrate yesterday and touched my face under my nose over 
my lip and now I have black spots that won't come off.  I have done this on my 
hands before but never on my face.  So far I have tried soaking a cloth in 
hydrogen peroxide hoping to bleach it with no luck, I even tried putting some 
gold chloride on it to see if I could tone it down, to no avail.  Any ideas?  
Make up only goes so far and lasts so long.

 

Regards,

 

Patsy

 

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net

 


This email is confidential and intended solely for the use of the Person(s) 
('the intended recipient') to whom it was addressed. Any views or opinions 
presented are solely those of the author. It may contain information that is 
privileged  confidential within the meaning of applicable law. Accordingly any 
dissemination, distribution, copying, or other use of this message, or any of 
its contents, by any person other than the intended recipient may constitute a 
breach of civil or criminal law and is strictly prohibited. If you are NOT the 
intended recipient please contact the sender and dispose of this e-mail as soon 
as possible.

 

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CONFIDENTIALITY NOTICE:
 The information in this e-mail may be confidential and/or
 privileged.  If you are not the intended recipient or an
 authorized representative of the intended recipient, you
 are hereby notified that any review, dissemination, or
 copying of this e-mail and its attachments, if any, or
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RE: [Histonet] help (UNCLASSIFIED)

[Gladney, Diane C Ms CIV USA MEDCOM MACH]

Classification:  UNCLASSIFIED 
Caveats: NONE

Patsy,

I have removed Silver Nitrate on my hands by first treating the stain with 
Iodine solution then flushing the area with Sodium Thiosulfate solution (5% ?) 
then washing my hands thoroughly with soap and water. It removed the Silver 
Nitrate. This was a trick that one of my instructors used while I was in 
school. She got the idea from one of the silver stains that we were doing 
(Reticulin, I think). I don't know if how it will work on your face or if it 
will irritate the delicate skin but I think that I would try it anyway just to 
get rid of the black mark. 

Thanks for the laugh and my heart goes out to you. 

Diane G.

Diane C. Gladney, HT (ASCP)
Supervisor, Anatomical Pathology 
Moncrief Army Community Hospital
Dept. of Pathology
4500 Stuart St.
FT. Jackson, SC  29207

Email:  diane.glad...@amedd.army.mil

Phone:  803-751- 2530 
FAX: 803-751-7829
DSN:    734-2530






-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg
Sent: Tuesday, March 23, 2010 12:00 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] help

After you stop laughing seriously I need some help here, apparently I got my
fingers in some silver nitrate yesterday and touched my face under my nose
over my lip and now I have black spots that won't come off.  I have done
this on my hands before but never on my face.  So far I have tried soaking a
cloth in hydrogen peroxide hoping to bleach it with no luck, I even tried
putting some gold chloride on it to see if I could tone it down, to no
avail.  Any ideas?  Make up only goes so far and lasts so long.

 

Regards,

 

Patsy

 

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net

 


This email is confidential and intended solely for the use of the Person(s)
('the intended recipient') to whom it was addressed. Any views or opinions
presented are solely those of the author. It may contain information that is
privileged  confidential within the meaning of applicable law. Accordingly
any dissemination, distribution, copying, or other use of this message, or
any of its contents, by any person other than the intended recipient may
constitute a breach of civil or criminal law and is strictly prohibited. If
you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.

 

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Classification:  UNCLASSIFIED 
Caveats: NONE


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RE: [Histonet] help

[Patsy Ruegg]

Thanks for all the ideas, the qtip with straight bleach worked but now I
have to treat the 3rd degree burns I have incurred thru all this.

Regards,

Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net
 

This email is confidential and intended solely for the use of the Person(s)
('the intended recipient') to whom it was addressed. Any views or opinions
presented are solely those of the author. It may contain information that is
privileged  confidential within the meaning of applicable law. Accordingly
any dissemination, distribution, copying, or other use of this message, or
any of its contents, by any person other than the intended recipient may
constitute a breach of civil or criminal law and is strictly prohibited. If
you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.


-Original Message-
From: Weems, Joyce [mailto:jwe...@sjha.org] 
Sent: Tuesday, March 23, 2010 10:08 AM
To: Patsy Ruegg; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] help

I think there is none... But thanks for the funny!! j 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg
Sent: Tuesday, March 23, 2010 12:00
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] help

After you stop laughing seriously I need some help here, apparently I got my
fingers in some silver nitrate yesterday and touched my face under my nose
over my lip and now I have black spots that won't come off.  I have done
this on my hands before but never on my face.  So far I have tried soaking a
cloth in hydrogen peroxide hoping to bleach it with no luck, I even tried
putting some gold chloride on it to see if I could tone it down, to no
avail.  Any ideas?  Make up only goes so far and lasts so long.

 

Regards,

 

Patsy

 

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net

 


This email is confidential and intended solely for the use of the Person(s)
('the intended recipient') to whom it was addressed. Any views or opinions
presented are solely those of the author. It may contain information that is
privileged  confidential within the meaning of applicable law. Accordingly
any dissemination, distribution, copying, or other use of this message, or
any of its contents, by any person other than the intended recipient may
constitute a breach of civil or criminal law and is strictly prohibited. If
you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.

 

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It may contain information that is privileged and 
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sender that you have received the message in 
error, then delete this message.




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RE: [Histonet] help

[Bell, Lynne]

Pretty funny stuff!!  I say to try the sodium thio - it removes unreduced 
silver.  Or you could grab a Brillo pad and scrub away!

This laugh was just what I needed today.  Thank you.

Lynne A. Bell, HT (ASCP)
Technical Specialist, Histology
Central Vermont Medical Center
130 Fisher Road
Barre, VT  05641
802-371-4923


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RE: [Histonet] help (UNCLASSIFIED)

[Maxim Peshkov]

Patsy:
Chemically, silver nitrate deposites can be
reduced onto the slides by 0.5% potassium ferricyanide.
I am not sure that it may work onto living skin.
Sincerely,
Maxim Peshkov,
Russia,
Taganrog.
   mailto:maxim...@mail.ru


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RE: [Histonet] help (UNCLASSIFIED)

[Patsy Ruegg]

Are you trying to kill me?

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net
 

This email is confidential and intended solely for the use of the Person(s)
('the intended recipient') to whom it was addressed. Any views or opinions
presented are solely those of the author. It may contain information that is
privileged  confidential within the meaning of applicable law. Accordingly
any dissemination, distribution, copying, or other use of this message, or
any of its contents, by any person other than the intended recipient may
constitute a breach of civil or criminal law and is strictly prohibited. If
you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.

-Original Message-
From: Maxim Peshkov [mailto:maxim...@mail.ru] 
Sent: Tuesday, March 23, 2010 11:43 AM
To: pru...@ihctech.net
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] help (UNCLASSIFIED)

Patsy:
Chemically, silver nitrate deposites can be
reduced onto the slides by 0.5% potassium ferricyanide.
I am not sure that it may work onto living skin.
Sincerely,
Maxim Peshkov,
Russia,
Taganrog.
   mailto:maxim...@mail.ru



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Re: [Histonet] help for silver

[John Kiernan]

Some 35 years ago, a person older than I told me the most effective way to get 
silver stains off skin was to rub the affected part with slightly dampened 
mercuric chloride powder. He may have been right; I never tried it! I've tried 
things like iodine-thiosulphate and Farmer's reducer (ferricyanide + 
thiosulphate mixture) but my experience is that they do very little. The 
blackened epidermis flakes off, usually in a couple of days.
 
John Kiernan
Anatomy, UWO
London, Canada.
= = =
- Original Message -
From: louise renton louise.ren...@gmail.com
Date: Tuesday, March 23, 2010 13:58
Subject: [Histonet] help for silver
To: Patsy Ruegg pru...@ihctech.net, Histonet@lists.utsouthwestern.edu

 I know we used Sodium thiosulphate and alcoholic iodine, but i cannot
 remember which ne first...I think the iodine??
 
 John Kiernan - help please??
 
 On Tue, Mar 23, 2010 at 5:59 PM, Patsy Ruegg 
 pru...@ihctech.net wrote:
 
   After you stop laughing seriously I need some help here, 
 apparently I got
  my
  fingers in some silver nitrate yesterday and touched my face 
 under my nose
  over my lip and now I have black spots that won't come 
 off.  I have done
  this on my hands before but never on my face.  So far I 
 have tried soaking
  a
  cloth in hydrogen peroxide hoping to bleach it with no luck, I 
 even tried
  putting some gold chloride on it to see if I could tone it 
 down, to no
  avail.  Any ideas?  Make up only goes so far and 
 lasts so long.
 
 
 
  Regards,
 
 
 
  Patsy
 
 
 
  Patsy Ruegg, HT(ASCP)QIHC
  IHCtech, LLC
  Fitzsimmons BioScience Park
  12635 Montview Blvd. Suite 215
  Aurora, CO 80010
  P-720-859-4060
  F-720-859-4110
  wk email pru...@ihctech.net
  web site www.ihctech.net
 
 
 
 
  This email is confidential and intended solely for the use of 
 the Person(s)
  ('the intended recipient') to whom it was addressed. Any views 
 or opinions
  presented are solely those of the author. It may contain 
 information that
  is
  privileged  confidential within the meaning of applicable 
 law. Accordingly
  any dissemination, distribution, copying, or other use of this 
 message, or
  any of its contents, by any person other than the intended 
 recipient may
  constitute a breach of civil or criminal law and is strictly 
 prohibited. If
  you are NOT the intended recipient please contact the sender 
 and dispose of
  this e-mail as soon as possible.
 
 
 
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 -- 
 Louise Renton
 Bone Research Unit
 University of the Witwatersrand
 Johannesburg
 South Africa
 +27 11 717 2298 (tel  fax)
 073 5574456 (emergencies only)
 There are nights when the wolves are silent and only the moon howls.
 George Carlin
 No trees were killed in the sending of this message.
 However, many electrons were terribly inconvenienced.
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Re: [Histonet] help

[Rene J Buesa]

Touch the affected areas with Lugol's solution. After that remove the Lugol 
with sodium thiosulfate.
René J.

--- On Tue, 3/23/10, Patsy Ruegg pru...@ihctech.net wrote:


From: Patsy Ruegg pru...@ihctech.net
Subject: [Histonet] help
To: histonet@lists.utsouthwestern.edu
Date: Tuesday, March 23, 2010, 11:59 AM


After you stop laughing seriously I need some help here, apparently I got my
fingers in some silver nitrate yesterday and touched my face under my nose
over my lip and now I have black spots that won't come off.  I have done
this on my hands before but never on my face.  So far I have tried soaking a
cloth in hydrogen peroxide hoping to bleach it with no luck, I even tried
putting some gold chloride on it to see if I could tone it down, to no
avail.  Any ideas?  Make up only goes so far and lasts so long.



Regards,



Patsy



Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net




This email is confidential and intended solely for the use of the Person(s)
('the intended recipient') to whom it was addressed. Any views or opinions
presented are solely those of the author. It may contain information that is
privileged  confidential within the meaning of applicable law. Accordingly
any dissemination, distribution, copying, or other use of this message, or
any of its contents, by any person other than the intended recipient may
constitute a breach of civil or criminal law and is strictly prohibited. If
you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.



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RE: [Histonet] help (UNCLASSIFIED)

[Kim . Donadio]

LOL!  I am so sorry Patsy I have been reading this unfortunate but 
extremely funny story and I just could not help myself any more. I know 
this has been such a pain in the hiney for you to have to deal with and I 
am sure all of us here sympathize. It's still funny though  you got to 
just laugh it off sometimes  :-) 

I sincerely hope you do not have 3rd degree burns. I have no idea what can 
take this off other than time. I think derm abrasion might have been the 
only thing that might  work. It's expensive though. 

And just to make you feel a little better. You are not the only person 
this has happened to, you're not alone. I think perhaps that's why many of 
us got such a laugh out of it.  or some of us could just want to pick 

Good luck getting that off! oh and just for fun, print this whole 
conversation off and when this is over for you. Go back and read it for 
fun  :o) 



Kim Donadio 
Pathology Supervisor
Baptist Hospital
1000 W Moreno St.
Pensacola FL 32501
Phone (850) 469-7718
Fax (850) 434-4996



Patsy Ruegg pru...@ihctech.net 
Sent by: histonet-boun...@lists.utsouthwestern.edu
03/23/2010 01:33 PM

To
'Maxim Peshkov' maxim...@mail.ru
cc
histonet@lists.utsouthwestern.edu
Subject
RE: [Histonet] help (UNCLASSIFIED)






Are you trying to kill me?

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pru...@ihctech.net
web site www.ihctech.net
 

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-Original Message-
From: Maxim Peshkov [mailto:maxim...@mail.ru] 
Sent: Tuesday, March 23, 2010 11:43 AM
To: pru...@ihctech.net
Cc: histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] help (UNCLASSIFIED)

Patsy:
Chemically, silver nitrate deposites can be
reduced onto the slides by 0.5% potassium ferricyanide.
I am not sure that it may work onto living skin.
Sincerely,
Maxim Peshkov,
Russia,
Taganrog.
   mailto:maxim...@mail.ru



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RE: [Histonet] Help...

[Liz Chlipala]

Fawn

CAP has specific quidelines, its right there in the questions, did you
do what that question is asking and have you documented what was done.
They tell you right in the check sheet what needs to be completed.  Test
validation must be performed on a minimum of 25 cases (they recommend
25-  100) and they tell you how to validate too.  There is also an
article that you can get on the CAP website on IHC standardization. I
have a pdf of it here if you need it.

LIz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Fawn
Bomar
Sent: Thursday, February 11, 2010 12:07 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Help...

Does anyone have a protocol or procedure for validating the Her2
antibody?  We are trying to get ready for a CAP inspection and I'm not
sure if what we did is ok or not.

Thank you in advance
Fawn
-
This electronic message may contain information that is 
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Re: [Histonet] Help...

[Rene J Buesa]

And what did you do?
René J.

--- On Thu, 2/11/10, Fawn Bomar fawn.bo...@halifaxregional.com wrote:


From: Fawn Bomar fawn.bo...@halifaxregional.com
Subject: [Histonet] Help...
To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu
Date: Thursday, February 11, 2010, 2:06 PM


Does anyone have a protocol or procedure for validating the Her2 antibody?  We 
are trying to get ready for a CAP inspection and I'm not sure if what we did is 
ok or not.

Thank you in advance
Fawn
-
This electronic message may contain information that is 
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in the message. 

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notify the sender immediately and delete the material from any 
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do not disclose its contents or take any action in reliance on
the information it contains. 

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RE: [Histonet] Help! Losing sections from Superfrost Plus Slides

[CHRISTIE GOWAN]

I would suggest you check the lot#. You may have received a bad lot and need to 
get them replaced by the vendor. 
 
 
 Hi all,
 
 We have suddenly started losing tissue sections from our Superfrost Plus 
 Slides. 
 Our students have been cutting fixed, frozen, cryosections (20um) and 
 thaw-mounting these onto Superfrost Plus slides.  We have suddenly started 
 losing lots of sections from these slides (again!!).  The tissue is small - 
 ie cross sections of frog aorta and longitudinal sections of nerves, so any 
 lose of adhesion results in total loss of the tissue.  The odd thing is that 
 she is not losing every section on every slide, but half to 3/4 of the 
 sections are falling off within the first rinse.  The sections are from the 
 same tissue block on the same slide while some falls and others don't.
 
 We are at a lose as to what we can do to rescue these sections.
 
 Does anyone know if there is any way to coat the slides in some solution with 
 the tissue on them to help improve the adhesion without losing the ability to 
 do immunofluorescence?
 
 Any further advice on cutting / drying protocols are welcomed. 
 This keeps happening and the inconsistency of it has us so frustrated with 
 this that we are thinking of going back to subbing our own slides.
 
 Thanks for your help!
 
 Angelina
 
 
 -- ~~o~~o~~o~~o~~o~~o~~o~~o~~o~~o
 
 Angelina Y. Fong, Ph.D.
 Department of Zoology
 Biological Sciences Building
 6270 University Boulevard
 University of British Columbia
 Vancouver, BC, V6T 1Z4
 Canada 
 Ph:  (604) 822-5799
 Fax: (604) 822-2416
 Email: f...@zoology.ubc.ca
 
 
 
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RE: [Histonet] HELP Starting a lab

[JR R]

Pretty good list.

Don't forget first aid kits and spill kits.

Jerry Ricks
Research Scientist
University of Washington
Department of Pathology

 Date: Mon, 20 Jul 2009 14:26:57 -0400
 From: ktut...@umm.edu
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] HELP Starting a lab
 
 I have a friend who is a Pathologist and starting a new lab. She sent me a 
 list of start up supplies and equipment that she needs to purchase. Can I get 
 some input on whats missing from her list, I've added a few things but Im 
 going blank. Its a hisology lab for a urology practice if that helps. Thank 
 you in advance!
 
 PLEASE VENDORS DONT CALL OR EMAL ME
 
 Equipment:
 Negative pressure hood for grossing of specimens (grossing station)
 Refrigerator w/freezer or separate freezer
 Cassette labeler
 Tissue processor
 Embedding Stations
 Microtomes
 Waterbaths,  1 per microtome
 Automatic HE stainer
 A scale and a mixing/heating plate
 Manual staining dishes, and slide holders that fit in them.
 Slide and block storage system
 
 Supplies:
 Safety equipment like goggles and sleeve covers, aprons, a flame cabinet for 
 flammable liquids 
 acid cabinet?
 Blades
 Forceps
 Rulers
 Hemostats
 Scissors
 Glassware like flasks, beakers, graduated cylinders, and a funnel.Pipettes 
 and tips.
 
 Reagents:
 Formalin
 70% etoh
 95% etoh
 100% etoh
 xylene 
 paraffin
 Hematoxylin stain
 Eosin stain
 
 
 Kimberly C. Tuttle  HT (ASCP)
 Pathology Biorepository and Research Core
 University of Maryland 
 Room NBW58, UMMC
 22 S. Greene St
 Baltimore, MD 21201
 (410) 328-5524
 (410) 328-5508 fax 
 Please consider the environment before printing this e-mail.
 
 
  
 
 This e-mail and any accompanying attachments may be privileged, confidential, 
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 confidentiality of this information. If the reader of this message is not the 
 intended recipient; you are prohibited from using, disclosing, reproducing or 
 distributing this information; you should immediately notify the sender by 
 telephone or e-mail and delete this e-mail.
 
 
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RE: [Histonet] Help With Hemo Fading

[Kemlo Rogerson]

I agree.. Also you're not storing them in sunlight are you? Silly question 
I know.



Kemlo Rogerson  
e-mail kemloroger...@nhs.net if not at work.
DD   01934 647057 or extension 3311 Mob 07749 754194; 
Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah Lehrer
This e-mail is confidential and privileged. If you are not the intended 
recipient please accept my apologies; please do not disclose, copy or 
distribute information in this e-mail or take any action in reliance on its 
contents: to do so is strictly prohibited and may be unlawful. Please inform me 
that this message has gone astray before deleting it. Thank you for your 
co-operation

 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: 02 July 2009 20:28
To: histonet@lists.utsouthwestern.edu; sr...@aol.com
Subject: Re: [Histonet] Help With Hemo Fading

The fading most probably is caused by acid in the permanent slide, probably 
because the sections were passed through the alcohols very quickly after the 
acid differentiation, or they stayed little time in tap water after 
differentiation or no bluing agent was used.
It is unlikely that the mounting medium is acidic, although that could also be 
the cause also. An acid environment over the cover slipped section is the most 
probable culprit for the henatoxylin fading.
Check the staining protocol.
René J.

--- On Thu, 7/2/09, sr...@aol.com sr...@aol.com wrote:


From: sr...@aol.com sr...@aol.com
Subject: [Histonet] Help With Hemo Fading
To: histonet@lists.utsouthwestern.edu
Date: Thursday, July 2, 2009, 2:42 PM



Hello everyone,

?

We are having problems with short-term hematoxylin fading and loss of detail. 
The pathologist is freaking out! I've seen hemo fade over a long period of time 
but not in a matter of a few months. Slides from one year ago are really bad.

?

I've been out of the business for a number of years and in the interim much has 
changed including reagents. These are GI tract biopsies processed by microwave. 

?

Any thoughts at all?

?

Thanks! Marg
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Re: [Histonet] Help With Hemo Fading

[louise renton]

Even long term exposure to indoor lighting (fluorescent type) will fade
sectiions. We work in a building where ther lights are permanently on. If we
want to keep our sections fresh and helathy, they get covered up as soon as
.

best regards

On Fri, Jul 3, 2009 at 9:09 AM, Kemlo Rogerson 
kemlo.roger...@waht.swest.nhs.uk wrote:

 I agree.. Also you're not storing them in sunlight are you? Silly
 question I know.



 Kemlo Rogerson
 e-mail kemloroger...@nhs.net if not at work.
 DD   01934 647057 or extension 3311 Mob 07749 754194;
 Embrace uncertainty. Hard problems rarely have easy solutions. --Jonah
 Lehrer
 This e-mail is confidential and privileged. If you are not the intended
 recipient please accept my apologies; please do not disclose, copy or
 distribute information in this e-mail or take any action in reliance on its
 contents: to do so is strictly prohibited and may be unlawful. Please inform
 me that this message has gone astray before deleting it. Thank you for your
 co-operation



 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
 Sent: 02 July 2009 20:28
 To: histonet@lists.utsouthwestern.edu; sr...@aol.com
 Subject: Re: [Histonet] Help With Hemo Fading

 The fading most probably is caused by acid in the permanent slide, probably
 because the sections were passed through the alcohols very quickly after the
 acid differentiation, or they stayed little time in tap water after
 differentiation or no bluing agent was used.
 It is unlikely that the mounting medium is acidic, although that could also
 be the cause also. An acid environment over the cover slipped section is the
 most probable culprit for the henatoxylin fading.
 Check the staining protocol.
 René J.

 --- On Thu, 7/2/09, sr...@aol.com sr...@aol.com wrote:


 From: sr...@aol.com sr...@aol.com
 Subject: [Histonet] Help With Hemo Fading
 To: histonet@lists.utsouthwestern.edu
 Date: Thursday, July 2, 2009, 2:42 PM



 Hello everyone,

 ?

 We are having problems with short-term hematoxylin fading and loss of
 detail. The pathologist is freaking out! I've seen hemo fade over a long
 period of time but not in a matter of a few months. Slides from one year ago
 are really bad.

 ?

 I've been out of the business for a number of years and in the interim much
 has changed including reagents. These are GI tract biopsies processed by
 microwave.

 ?

 Any thoughts at all?

 ?

 Thanks! Marg
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South Africa
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No trees were killed in the sending of this message.
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Re: [Histonet] Help With Hemo Fading

[Rene J Buesa]

The fading most probably is caused by acid in the permanent slide, probably 
because the sections were passed through the alcohols very quickly after the 
acid differentiation, or they stayed little time in tap water after 
differentiation or no bluing agent was used.
It is unlikely that the mounting medium is acidic, although that could also be 
the cause also. An acid environment over the cover slipped section is the most 
probable culprit for the henatoxylin fading.
Check the staining protocol.
René J.

--- On Thu, 7/2/09, sr...@aol.com sr...@aol.com wrote:


From: sr...@aol.com sr...@aol.com
Subject: [Histonet] Help With Hemo Fading
To: histonet@lists.utsouthwestern.edu
Date: Thursday, July 2, 2009, 2:42 PM



Hello everyone,

?

We are having problems with short-term hematoxylin fading and loss of detail. 
The pathologist is freaking out! I've seen hemo fade over a long period of time 
but not in a matter of a few months. Slides from one year ago are really bad.

?

I've been out of the business for a number of years and in the interim much has 
changed including reagents. These are GI tract biopsies processed by microwave. 

?

Any thoughts at all?

?

Thanks! Marg
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RE: [Histonet] Help With Hemo Fading

[Monson, Frederick]

 messy that it had been recommended that I submit to a bone marrow exam 
to determine my proximity to death.  I quietly demanded to know how the 
differential was done.  The answer?  The Coulter counter!  I quietly agreed to 
a bone marrow exam AFTER a manual differential was performed.  Later, the 
hematologist only wanted to know how I knew.  I told her that while working in 
a path lab at night during college (I know I wasn't 'certified'!!!), I had 
compared myself to the 'Counter' and I was frankly more consistent.  I admitted 
that my test was not statistical, but I had real confidence in the capacity of 
the eye and little in the photometer looking thru glass.

Machines may be necessary to do the work, but they do not clean themselves and 
they do not cry out when the 95% ethanol has become 87%.  With Coplin jars, 
variables were easy to manage.  They were more difficult the more we tried to 
do multiples of specimens.  This should be obvious in spite of the fact that 
automation is practical and manual is not - even though sometimes necessary.

When I started in histology, I began at the beginning of the Paraplast 
revolution, yet there were still those who formulated their own paraffin for 
the part of the planet in which they worked.

Sorry for the late-night babble, but I have vented stored up compulsion.

Cheers,

Fred Monson

P.S.  

Those who are required to volunteer do NOT.  Those who require others to 
volunteer fail.

Every tax is a means by which a government forces a citizen to pay for the 
vacations of those who work for the government.

When the USA has a public health plan, every union will be forced to accept it 
by the employer, and all those who want something better will be required to 
pay for the full load.  Government plan - 'free', all others - cost extra.  
That's competition?  Why won't the insurance companies want to give 'free' 
public health care to their employees?  Their own actuaries will show them the 
way.
 
Three of four wars prosecuted by the United States during the 20th century were 
begun under democratically elected Presidents.  

One was fought entirely by volunteers.  

During only two of the four have a majority of citizens, at one time or 
another, wished the U.S.A. to lose(???).  

Three cheers for the Draft!  

Beware the resolve of an elected President not to lose.

Frederick C. Monson, PhD
Technical Director, CMIRT
West Chester University
West Chester, PA, 19383
610-738-0437
http://cmirt.wcupa.edu


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu on behalf of Rene J Buesa
Sent: Thu 7/2/2009 3:27 PM
To: histonet@lists.utsouthwestern.edu; sr...@aol.com
Subject: Re: [Histonet] Help With Hemo Fading
 
The fading most probably is caused by acid in the permanent slide, probably 
because the sections were passed through the alcohols very quickly after the 
acid differentiation, or they stayed little time in tap water after 
differentiation or no bluing agent was used.
It is unlikely that the mounting medium is acidic, although that could also be 
the cause also. An acid environment over the cover slipped section is the most 
probable culprit for the henatoxylin fading.
Check the staining protocol.
René J.

--- On Thu, 7/2/09, sr...@aol.com sr...@aol.com wrote:


From: sr...@aol.com sr...@aol.com
Subject: [Histonet] Help With Hemo Fading
To: histonet@lists.utsouthwestern.edu
Date: Thursday, July 2, 2009, 2:42 PM



Hello everyone,

?

We are having problems with short-term hematoxylin fading and loss of detail. 
The pathologist is freaking out! I've seen hemo fade over a long period of time 
but not in a matter of a few months. Slides from one year ago are really bad.

?

I've been out of the business for a number of years and in the interim much has 
changed including reagents. These are GI tract biopsies processed by microwave. 

?

Any thoughts at all?

?

Thanks! Marg
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RE: [Histonet] help needed to find parts for tissue processor

[Paul Firnschild]

Leroy,

You could also have a electronics technician troubleshoot and repair your
screens.  You know. Transistors, diodes, triacs, capacitors which are all
available online. Hardly anyone even thinks of that option anymore.  Let me
know if I can help you.

Paul
 
Paul M. Firnschild
QA Support Services, Inc.
404.291.3715
 

-Original Message-
From: Leroy Brown [mailto:rhbro...@histocs.com] 
Sent: Wednesday, May 27, 2009 12:24 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] help needed to find parts for tissue processor

Hi ,  I have two MVP-1 tissue processor  that are still in great shape
except that both have lost the display  screens.   Other than that they are
wonderful tissue  processors.I am told by the folks that know this
machine that I can swap out these screens from any working model from  any
of the following processors.   The RMC models,  the MVP-1  or MVP-2 the
older 1530 , the Renaissance processor.   They are  all similar since this
machine changed hands so many times.  It was a  Fisher Scientific,  then
Instrumentation Lab,  then Vantana  and so on.   The problem is that the
parts are not available any  longer.My only hope if I want to continue
using these  processors is to find a parts machine or one that is sitting in
the basement  of someones hospital or lab perhaps for years.  So, if you
have one or  know where to find one I would very much like to hear from you.
I bought these two processors about 2 years ago from a  government auction
still new in the ori
ginal crate  unopened.I really would like to get them  up and running
again.   

Thanks  

LeRoy Brown HT(ASCP) HTL  

www.histocs.com  

email to me directly at lhb...@comcast.net or call me at  360-966-7300



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Re: [Histonet] help

[Rene J Buesa]

Your advise to her is correct: probably the sections are not destained 
completely.
Any section stained with the Ziehl-Nielsen Carbol Fucsin has to be treated with 
acid alcohol until no more red color leaches from the section no matter how 
much time that takes. No more red coming out of the section is the end point of 
the destaining.
René J. 

--- On Sun, 4/26/09, Dertien, Janet janet.dert...@ttuhsc.edu wrote:

From: Dertien, Janet janet.dert...@ttuhsc.edu
Subject: [Histonet] help
To: histonet@lists.utsouthwestern.edu
Date: Sunday, April 26, 2009, 3:16 PM

Hello, I have a friend who is having trouble with ziehl-neesen stainin. Her
sections appear red (with no blue). I have a feeling she may not be destaining
adequately. Any suggestions?


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu on behalf of
histonet-requ...@lists.utsouthwestern.edu
Sent: Sun 4/26/2009 12:05 PM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 65, Issue 44
 
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Today's Topics:

   1. paraffin embedding (Marc Shaeffer)
   2. Long term storage for IHC? (Mikael Niku)
   3. Re: Long term storage for IHC? ( TF )
   4. AW: [Histonet] Long term storage for IHC? (Gudrun Lang)
   5. Long term storage for IHC ? (Rene J Buesa)


--

Message: 1
Date: Sat, 25 Apr 2009 10:15:50 -0700
From: Marc Shaeffer mshaef...@cox.net
Subject: [Histonet] paraffin embedding
To: histonet@lists.utsouthwestern.edu
Message-ID: b7392e4768d3431d997fb5694d0c7...@youro0kwkw9jwc
Content-Type: text/plain;   charset=us-ascii

Don't forget to put the plastic cassette on top the mold prior to filling
and solidifying the paraffin.



--

Message: 2
Date: Sat, 25 Apr 2009 23:24:56 +0300
From: Mikael Niku mikael.n...@helsinki.fi
Subject: [Histonet] Long term storage for IHC?
To: histonet@lists.utsouthwestern.edu
Message-ID: 49f37198.5030...@helsinki.fi
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

Dear Histonetters,

what do you think is the best way to store formalin-fixed tissues for 
immunohistochemistry, long-term (several years)?

We are currently fixing overnight, then rinsing in buffer and storing in 
70% or 100% ethanol, +4C or -20C. However, I don't really KNOW what 
happens here and how the tissues will work after the years. I guess 
basically the formalin fixation will be at least somewhat reversed and 
after some time the tissues will be more like ethanol fixed. But are 
there better options? Long term storage in formalin will make IHC 
difficult. And this is about large numbers of tissue samples, only some 
of which will be actually used later, so we wouldn't like to process all 
of them to paraffin either.

With best regards,
Mikael Niku
University of Helsinki, Finland



--

Message: 3
Date: Sun, 26 Apr 2009 13:19:53 +0800
From:  TF  ti...@foxmail.com
Subject: Re: [Histonet] Long term storage for IHC?
To:  Mikael Niku  mikael.n...@helsinki.fi,  histonet

histonet@lists.utsouthwestern.edu
Message-ID: 200904261319483656...@foxmail.com
Content-Type: text/plain;   charset=utf-8

Hi, i embeded all of them in OCT and put them under -20.
To prevent water vapor leakage, use some parafilm to pack the OCT block or seal
it.
We tested this in samples harvested 5 years ago (2004-2009).

2009-04-26 



TF 



å�'件人ï¼s Mikael Niku 
å�'é?�æ-¶é-´ï¼s 2009-04-26  04:31:05 
æ¶ä»¶äººï¼s histonet 
æSé?�ï¼s 
主é¢~ï¼s [Histonet] Long term storage for IHC? 
 
Dear Histonetters,
what do you think is the best way to store formalin-fixed tissues for 
immunohistochemistry, long-term (several years)?
We are currently fixing overnight, then rinsing in buffer and storing in 
70% or 100% ethanol, +4C or -20C. However, I don't really KNOW what 
happens here and how the tissues will work after the years. I guess 
basically the formalin fixation will be at least somewhat reversed and 
after some time the tissues will be more like ethanol fixed. But are 
there better options? Long term storage in formalin will make IHC 
difficult. And this is about large numbers of tissue samples, only some 
of which will be actually used later, so we wouldn't like to process all 
of them to paraffin either.
With best regards,
Mikael Niku
University of Helsinki, Finland
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Re: [Histonet] Help with protocols

[Histonet Alias]

If the CC1 standard 32 with amp, 42 degrees does not work then try using
protease 1 4-6 minutes on it.

On Fri, Nov 28, 2008 at 12:35 PM, Lena Spencer [EMAIL PROTECTED]wrote:

 Hi All:

 I have been working up the PMS2 and MLH2 mismatched repair genes and I am
 not satisfied with my results.  I am using the Cell Marque antibodies and
 the Ventana XT using CC1, mild, standard and I have tried the antibodies
 with block and amplification, and antibody times from 32 min- 1 hour and
 the
 stain is not satisfactory.  Anyone who has these antibodies in their lab
 and
 would like to share your  protocols,  I would be grateful for your help.
 The MSH2 and MSH6 are working great.

 Lena Spencer

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