[Histonet] processing tissue with silicone mesh in it

[pruegg]

Does anyone have any experience with processing tissues with silicone mesh
in them so that the silicone is not dissolved by xylene and we also assume
that it might be dissolved by MMA or GMA polymers since they are strong
solvents in their own right?

 

Thank you,

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 

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[Histonet] RE: IHC on animals

[pruegg]

Lester,

Probably not in most cases but it depends upon what the antibody is, whether
as a an antihuman ab it also cross reacts to the species you are interested
in and what detection reagents you are using.
For instance if you have determined a rabbit anti human antibody also cross
reacts in mouse tissue, you could use a detection such as for the Leica Bond
by leaving out the link (they call it the post primary but it is rabbit anti
mouse IgG to link mouse antibodies to the goat anti rabbit labeled polymer
second step), since your ab is rabbit it would link directly to the anti rab
labeled polymer and the anti mouse link which would bind non specifically to
the mouse endogenous Ig can be avoided. 
You would have to do a lot of research to first determine if the anti human
rabbit antibody you want to use (it should not be made in a mouse) cross
reacts to mouse tissue.  The species you are wanting to label is also very
important, this system would only work for anti human rab abs that also
react in mouse, rat or any other species except rabbit and goat because
those are the species the detection is made in or against.
We have research versions of the Leica Bond instruments which allows us to
manipulate the instrument and detection for these purposes including
replacing the rab anti ms link with another link, I am not sure if the pure
clinical versions of the Bond allow this.
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 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth
Chlipala
Sent: Wednesday, March 26, 2014 2:20 PM
To: Lester Raff MD; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: IHC on animals

Lester

I would not, first of all not all antibodies that work in human will cross
react with a particular species of animal.  Another thing to consider is
that most human based detection systems consist of dual link reagents,
meaning that they work on both mouse and rabbit primaries these detection
system are not ideal for working with animals and depending upon the species
that you are working with may cause some problems in with background
staining due to the secondary antibody or polymer binding to endogenous IgG
in the samples.  You can initially pilot your protocol on a particular
species to see what happens but I would not take the chance and run a bunch
of slides thinking that the protocol that you have in place will work in the
particular species of animal you need to stain.

Good Luck

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 Lester Raff
MD
Sent: Wednesday, March 26, 2014 2:03 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC on animals

Hi:

 

Can IHC validated for humans be performed on animal tissues without any
protocol changes?

 

Thanks,

 

Lester J. Raff, MD MBA

UroPartners

Medical Director Of Laboratory

2225 Enterprise Dr. Suite 2511

Westchester, Il 60154

Tel: 708-486-0076

Fax: 708-492-0203

 

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[Histonet] RE: what is a level

[pruegg]

It can be confusing because people use different words to describe, we call
picking up every section "serial sectioning" and the distance between
sections for levels is determined by the requestor in our case, so they
might ask for six more levels to be cut with 50 microns  thrown out between
each level.  We leave that up to our customers to choose because they pay by
the slide.

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 



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Terri Braud
Sent: Tuesday, March 18, 2014 12:03 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: what is a level

Our 2 cents: What is a level?  A level is how you've defined level in your
SOP.  In this lab, we use the words level and recut interchangeably. We
define a level as 40 microns deeper than the previous section.  If the
pathologist wants to see every section of tissue cut without skipping any
tissue, then they order "sequential"
sections.  If we get orders to go into a block a second time, call it recut
or level, then the 1st slide is always the first full face section obtained.
If the tech senses that the orders will exhaust the tissue in the block,
then they communicate that to the pathologist before cutting.
If the pathologist orders the block to be cut through, we will ask them how
many slides they want and space them accordingly. Its not really that
complicated and it works for us.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Holy Redeemer Hospital Laboratory
1648 Huntingdon Pike
Meadowbrook, PA 19046
Ph: 215-938-3676
Fax: 215-938-3874

***

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[Histonet] Poster at USCAP

[pruegg]

Dear Colleagues,

 

If you are attending USCAP and get a chance take a look at this poster.

 

A couple of years ago I was involved in an IHC HIER study to compare work
done near sea level to work done at an altitude above 5K feet, it is the
first work of this kind since I wrote an article about HER2neu at altitude
in 2001, as far as we know.

 

This study will be presented as a poster at USCAP in San Diego, March 3rd #
2108.  This study shows IHC on the same 10 antibodies and same tissue,
comparing HIER at different temperatures both at low altitude (work done at
Emory U in Atlanta, Georgia with Dr. Cohen) and at an altitude above 5K feet
(work done at IHCtech in Aurora, Colorado with Patsy Ruegg).

 

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 

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[Histonet] RE: Mouse F4/80 antibody

[pruegg]

Rat anti ms F4/80 is used on mouse tissue ffpe for macrophages not anti
human cd68.

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 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Anna Hughes
Sent: Wednesday, February 12, 2014 9:14 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Mouse F4/80 antibody

Hi Everyone!
I was wondering if any of you have a favorite CD68 macrophage antibody that
you have used on mouse tissue and is especially reliable.  This is a target
that has been notorious in our lab and I was wondering if there was a really
good one to use.

Thanks in advance for your help!

Anna Hughes
Anna C. Hughes, BBA, BS, HTLCM
anna.c.hug...@gsk.com


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RE: [IHCRG] RE: [Histonet] Mouse GranzymeB

[pruegg]

I agree with what Liz has said.  Usually when I encounter undesirable bg
staining especially if it is nuclear it most often points to over retrieval
of tissues not adequately fixed.  Besides using less harsh retrieval such as
low ph HIER, EIER, or no IER Loui Harkin is a big believer of prefixing
questionably fixed tissue sections after depar in formalin before HIER and
that has worked for me in the past as well.  I almost routinely use serum
free protein block (casein) and in difficult cases replace that with 20%
serum from the species the secondary is made in.  Dako anti rab Envision+ LP
and Leica's anti rab Power Vision LP are my go too reagents for using rab
antibodies on ffpe mouse tissue.  Leica Bonds detection system can be used
for this as well if u skip the post primary which is rabbit anti mouse and
just go direction to the goat anti rab labeled polymer detection.

Good luck,

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 



 



-Original Message-
From: ih...@googlegroups.com [mailto:ih...@googlegroups.com] On Behalf Of
Elizabeth Chlipala
Sent: Tuesday, January 28, 2014 12:26 PM
To: pru...@ihctech.net; 'Amos Brooks'; histonet@lists.utsouthwestern.edu;
'ihcrg Group, (E-mail)'
Subject: RE: [IHCRG] RE: [Histonet] Mouse GranzymeB

Patsy and Amos

I can appreciate your comment but we have been using Dako's Envision+ Rabbit
on mouse tissue for years now and have never seen any cross reactivity to
mouse tissue, it's pretty much our go to detection system for rabbit
antibodies (both monoclonal and polyclonal) on mouse tissue.  We have
however seen cross reactivity to porcine with Dako's envision +mouse and
other companies anti-mouse polymer reagents.   My suggestion if you have not
already done this is to try to use some other retrieval methods such as pH6
or enzyme (proteinase K or pepsin or even no retrieval).  Tissue fixation is
important along with time an temp of retrieval - under fixation or over
retrieval or a combination of both can cause problems.  If we find we are
getting some background we will run some more intense blocking - such as the
following:

1. commercially available superblock - these can be competitive and may
potentially cause decreased sensitivity so you will need to watch out on
these 2.  Normal serum - we normally use a commercial serum free protein
block but on occasions we will use up to a 20% normal serum block for 30 to
60 minutes, if you want to continue to use the envision+ rabbit your choice
would be 20% normal goat serum 3. Saturated Casein - we'll make this up in
house

All are worth a try, good luck

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

Ship to Address:

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

-Original Message-
From: ih...@googlegroups.com [mailto:ih...@googlegroups.com] On Behalf Of
pru...@ihctech.net
Sent: Tuesday, January 28, 2014 12:11 PM
To: 'Amos Brooks'; histonet@lists.utsouthwestern.edu; 'ihcrg Group,
(E-mail)'
Subject: [IHCRG] RE: [Histonet] Mouse GranzymeB

Amos u might have to get an anti rabbit link that has been mouse absorbed.
Dako's anti rab envision is made in a goat but it is not mouse absorbed.
Southern Biotec or Jackson Labs are really good places to get absorbed
links.  Still no reliable cd4 and cd8 for ffpe mouse tissue as far as I
know, I have tried many that claim they work but have not consistently
worked in my hands, we still do that with frozen mouse tissue not aldehyde
fixed.  Are u doing HIER with the EDTA ph9?  In my experience especially
with the higher ph u can over retrieve and cause undesirable nuclear
staining.

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
rueggihcconsultin...@outlook.com



 

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: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amos Brooks
Sent: Friday, January 24, 2014 10:09 AM
To: histonet@lists.utsouthwestern.edu; ihcrg Group, (E-mail)
Subject: [Histonet] Mouse GranzymeB

Ok Musketeers,
 I am trying t

RE: [Histonet] Mouse GranzymeB

[pruegg]

Amos u might have to get an anti rabbit link that has been mouse absorbed.
Dako's anti rab envision is made in a goat but it is not mouse absorbed.
Southern Biotec or Jackson Labs are really good places to get absorbed
links.  Still no reliable cd4 and cd8 for ffpe mouse tissue as far as I
know, I have tried many that claim they work but have not consistently
worked in my hands, we still do that with frozen mouse tissue not aldehyde
fixed.  Are u doing HIER with the EDTA ph9?  In my experience especially
with the higher ph u can over retrieve and cause undesirable nuclear
staining.

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 
rueggihcconsultin...@outlook.com



 

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: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amos Brooks
Sent: Friday, January 24, 2014 10:09 AM
To: histonet@lists.utsouthwestern.edu; ihcrg Group, (E-mail)
Subject: [Histonet] Mouse GranzymeB

Ok Musketeers,
 I am trying to detect cytotoxic T-Cells in formalin fixed paraffin
embedded mouse liver. GranzymeB should do the trick. I have a rabbit anti
mouse (and human and rat) GranzymeB from abCam (ab53097). The spleen that I
ran as a control with it works fine. Nice T-cells and no signifigant
background staining. The liver on the others hand has some lymphocyte
staining, presumably cytotoxic T-cells as expected,  but it looks like every
hepatocyte nuclei are also picking it up.  Grrr,  right?
 If this antibody had been raised in mouse,  I wouldn't be surprised.
Also,  if the spleen looked similar, I wouldn't be surprised. Do any of you
have any ideas ab pi ut what this might be? Alternatively, does anyone have
any ideas about another antibody that would work here?  CD4 or CD8 would be
great here,  but its FFPE mouse,  so sadly that's out (more grumbling).
For the completionists out there,  I used EDTA pH9,  peroxide block and
detected the antibody (diluted 1:800) with Envision (rabbit) and DAB from
Dako.

Any suggestions would be vastly appreciated,

Happy Friday,
Amos
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RE: [Histonet] IHC-start-up

[pruegg]

We use the Leica platforms and just got a Leica RX which is for animal research 
and ISH, we love the openness of it.

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 
rueggihcconsultin...@outlook.com



 

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: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mike Thompson
Sent: Thursday, January 23, 2014 11:10 AM
To: Colleen Forster
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC-start-up

Diagnostic Biosystems has the same platform with tech support that is 
outstanding!

Michael O. Thompson
Director of Sales
Diagnostic BioSystems
Phone: 1-888-896-3350
Mobile: 412-860-1288
Office Fax: 412-727-6080

"IHC Made Affordable"
 www.dbiosys.com

Colleen Forster  wrote:

>I totally agree with William. I do animal IHC and I am, using the 
>Biocare Nemesis, which be the same as the Dako platform. Completely 
>open so you can do what you need to at a cost you can afford
>
>Colleen Forster HT(ASCP)QIHC
>u of MN
>
>
>
>On 1/23/2014 11:44 AM, Will Chappell wrote:
>> Doing ihc on animal tissue is very tricky, if for no other reason than you 
>> are trying to standardize an antibody across multiple species. You will need 
>> to use every trick in the book and invent some of your own to get 
>> reproducible results.
>>
>> I would steer clear of any platforms that do not let you customize 
>> everything. I would suggest an OLD Dako or biocare's IntelliPath. Both have 
>> their quirks but they have the openness you require.
>>
>> William Chappell
>>
>> Sent from my iPhone
>>
>>> On Jan 23, 2014, at 11:49 AM, Erin Sarricks  wrote:
>>>
>>> Hi all-
>>>
>>> Our histopathology lab is looking to set up an IHC component to 
>>> service the request of our clients.  All work is on animal tissue.  We will 
>>> probably run about 1,000 IHC slides the first year and hope to increase the 
>>> workload each year.  Some  stains we would run would include CD4, CD8, 
>>> Ki-67 and TUNEL. We looked at the Ventana Discovery XT and appeared to be a 
>>> good fit for our needs.  Does anyone have any other advice on other 
>>> machines we should look into?  Any information or advice would be greatly 
>>> appreciated. Thank you!
>>>
>>> Regards,
>>>
>>> Erin Sarricks, HT (ASCP)
>>> ___
>>> Histonet mailing list
>>> Histonet@lists.utsouthwestern.edu
>>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>> ___
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
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RE: [Histonet] OT-Retirement

[pruegg]

Very interesting post and life Ray, you should be proud of your accomplishments 
and contributions.  I agree and and share your concerns about our inadequate 
STEM training for our up coming generations.  Carl Sagan once said "our 
politicians and many inadequately trained teachers have a vested interest in 
not wanting the masses to think critically", because if they do they start 
asking questions the politicians and unfortunately many of the teachers feel 
uncomfortable answering because they most likely do not know the answers. It is 
too bad they feel threatened by questions they don't know the answers to rather 
than inspired to discovery the answers with the questioners.  We all know how 
politicians and too many teachers want us to think they are "all knowing", it 
is the bases of their power over us.  I believe this to be true and it really 
disturbs me.  I to am now retired since I sold my lab business a year and half 
ago.  I do try to do my part locally by serving on some boards at high schools 
promoting science education, especially laboratory science.  Maybe if we work 
together we can make a difference in our future generations, I want them to 
crave the STEM knowledge just like I did.  Let me know how I can help the cause 
to instill the passion for STEM knowledge so needed in our future generations.
 
Best regards,
 
Patsy Ruegg
 
- Original Message - Subject: [Histonet] OT-Retirement
From: koelli...@comcast.net
Date: 1/11/14 1:03 pm
To: histonet@lists.utsouthwestern.edu

Hello all out there, 
   
 This is regarding: Ray Koelling; currently from just north of the Seattle, WA 
area.  If you and I have connected in some way over the last 47 years, the 
following is a message concerning my (semi)-retirement that I hope you will 
enjoy and anyone with little or no interest in such writing or that knows me 
not, can easily use the delete button right now to save themselves a few 
minutes of OT reading. 
   
 It is time in life to shift a bit of focus after this long and amazing 
anatomic pathology journey.  There was the high school summer job in the mid 
60's at a St. Louis histopath lab changing a Lipshaw linear, open air tissue 
processor (we used dioxane both miscible with aqueous solutions and paraffin 
and NOT the dioxin of Agent Orange infamy) along with folding A LOT of paper 
boats for embedding and making 10% formalin from 55 gallon drums of 37% 
formaldehyde solution.  Somehow my brain has survived relatively unscathed.  
Some may dispute that last sentence.  Working at Jackson Memorial Hospital in 
South Florida with the great Dr. Azorides Morales and Dr. Mehred Nadji 
(actually Nadji was a resident when I was there and I have a treasured picture 
of him kicked back at a BBQ at my house wearing his famous sandals).  To 
learning more advanced immunology, histology and a lot more of 
cellular/molecular techniques along with the ability to critically think during 
the 5 years (of both unbelievably positive heaven and unrelenting, unforgiving 
hell) of graduate school.  To the biotechnology world and working on such drugs 
as etanercept, panitumumab, denosumab and (still in testing) TSLP, a compound 
that I had actually worked on in graduate school but when it was the newly 
discovered murine form TSLP and then also 50 other discovery molecules that all 
saw their shelving at various stages of development as failed candidates to 
progress in a particular pathway.  To then being able to work with Dr. Allen 
Gown in his fantastic lab in the Seattle area.  So if we have run into one 
another, in person, on-line, at a meeting or anywhere in the last nearly half 
century, hope you are doing well and are keeping safe and I wish you the best. 
   
 A part of my time now is going to be spent on some K-12 education projects.  
Organizing and helping at 2 different school districts for district-wide 
science fairs and STEM (science, technology, engineering, math) career 
festivals.  I am helping at the annual biotechnology fair in the Puget Sound 
Region for 300-400 high schoolers.  And am on Board of the Washington State 
Science and Engineering Fair that is the entrance point for this state into the 
big International Intel Science Fair.  Why?  It is no new, great news flash at 
all that the US is sinking further behind many countries of the world in math 
and science education.  And that is to the severe, possibly life-long detriment 
of kids now who will be less able to compete, as adults, with a global economy, 
jobs and society of the 21st century and which will frankly revolve a lot 
around STEM issues whether you like it or not.  The world is simply not now, 
nor will ever be again, as it was with me holding a 4-year degree in 
biology/chemistry in 1973 and at that time having virtually unlimited access to 
whatever I wanted to do. 
   
 Then for those kids K-12 who don't like math and science at all and don't want 
to be in STEM or any kind of STEM career I have offered

RE: AW: [Histonet] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde

[pruegg]

i would think u are correct in advising formic acid decal and then processing 
into paraffin for the best protection of the trap enzyme, immunoreactivity, 
etc.  A couple of weeks in formalin should be fine.  Paraformaldehyde show be 
the same as formalin.  I do know a way to restore the enzyme activity for TRAP 
that may have been lost so if u need that let me know.
 
- Original Message - Subject: AW: [Histonet] Bone samples 
long-term storage in 10% formalin or 4% paraformaldehyde
From: "Gudrun Lang" 
Date: 12/5/13 11:42 am
To: "'Orla M Gallagher'" 
Cc: histonet@lists.utsouthwestern.edu

Paraformaldehyd is formaldehyd in solid form. Formalin is the aequous
 solution of formaldehyd. 
 So the main characteristics are the same.
 
 Gudrun Lang
 
 -Ursprüngliche Nachricht-
 Von: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Orla M
 Gallagher
 Gesendet: Donnerstag, 05. Dezember 2013 19:31
 An: histonet@lists.utsouthwestern.edu
 Betreff: [Histonet] Bone samples long-term storage in 10% formalin or 4%
 paraformaldehyde
 
 Dear Histonetters,
 
 What is your opinion on storing bone samples long-term (more than a couple
 of weeks) in 10% formalin? As I was taught, best practice has always been to
 fix only as long as necessary, depending on the size of the sample, then
 decalcify and process to wax, and I always stress this to everyone I advise.
 
 However, research colleagues sometimes wish to do histology on bone samples
 that have been stored for months ..or even years! As the formalin pH becomes
 more acidic, there is formalin pigment and the immunoreactivity and TRAP
 enzyme activity is diminished or destroyed during long fixation, is there
 any way of minimising this e.g. has anyone tried regularly replacing the old
 formalin with fresh buffered formalin, or storing formalin-fixed bones in
 any other medium? I'm also interested in how best to fix in 4%
 paraformaldehyde and whether the problems are the same with long-term
 storage.
 
 Thanks for your comments.
 
 All the best,
 Orla
 
 --
 **
 Ms. Orla Gallagher
 Bone Analysis Laboratory
 Mellanby Centre for Bone Research
 Department of Human Metabolism
 D Floor Medical School
 University of Sheffield
 Beech Hill Road
 Sheffield
 S10 2RX
 UK
 
 Website: http://mellanbycentre.dept.shef.ac.uk
 
 Tel: 0044114-2713337 (office)
 0044114-2713174 (lab)
 E-Mail: o.m.gallag...@sheffield.ac.uk
 
 
 *STOP*: Do you really need to print this e-mail?
 
 *BE GREEN:* Keep it on the screen.
 
 
 *Times Higher Education University of the Year*
 
 
 
 Data protection and confidentiality:
 The information contained in this message or any appended documents may be
 privileged and confidential and is intended for the exclusive use of the
 addressee(s). If you are not the addressee, any disclosure, reproduction,
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 please contact the sender immediately and permanently delete/destroy what
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RE: [Histonet] Faxitron Xray digital imaging

[pruegg]

Years ago/maybe more than 25 we used a faxitron xray machine to measure bone
calcification on research samples and samples being decaled, it was the best
thing since sliced bread, had to go to the radiology dept to use it and was
told they were dying off the market, loved it, so easy to use and so useful.

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 
rueggihcconsultin...@outlook.com



 

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('the intended recipient') to whom it was addressed. Any views or opinions
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-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of WILLIAM
DESALVO
Sent: Tuesday, November 05, 2013 9:47 AM
To: Scott, Allison D; histonet
Subject: RE: [Histonet] Faxitron Xray digital imaging

We have used the Faxitron instrument for the past 3 years at multiple sites.
Faxitron creates a digital image of a specimen quickly and safely and a t
multiple magnifications. The instrument is placed in either Surgery or
Surgical Pathology (SP) and we use it primarily on breast cases to assist in
orientation and gross dissection, identifying radioactive seeds or locating
micro calcification. The instrument is easy to use, safe and you can connect
to multiple LIS systems and a Hospital PACS. There is no code to charge for
use when used in SP, only when used by Radiology or Surgery.
 
There is only one company that manufactures and sells the instrument:
Faxitron Bioptics, LLC
Tucson, AZ
877-910-0030
 
The are willing to bring a unit to your site for demonstration and you may
be able to talk them into leaving for a evaluation.
William DeSalvo, BS HTL(ASCP)

 
> From: allison.sc...@harrishealth.org
> To: histonet@lists.utsouthwestern.edu
> Date: Tue, 5 Nov 2013 16:00:11 +
> Subject: [Histonet] Faxitron Xray digital imaging
> 
> Hello to all in histoland.  Our chief pathologist is interested in getting
a faxitron xray digital imaging machine.  Is anyone out there using this
technology and can you suggest a company in which to order one from.  Any
help in this will be greatly appreciated.
> 
> 
> 
> 
> Allison Scott HT(ASCP)
> Supervisor, Histology Lab
> LBJ Hospital
> Harris Health System
> Office: 713-566-2148
> Lab: 713-566-5287
> 
> 
> CONFIDENTIALITY NOTICE:
> If you have received this e-mail in error, please immediately notify 
> the sender by return e-mail and delete this e-mail and any attachments 
> from your computer system.
> 
> To the extent the information in this e-mail and any attachments 
> contain protected health information as defined by the Health 
> Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 
> 104-191; 45 CFR Parts 160 and 164; or Chapter 181, Texas Health and 
> Safety Code, it is confidential and/or privileged.  This e-mail may 
> also be confidential and/or privileged under Texas law.  The e-mail is 
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RE: [Histonet] CD 34 antibody abcam (ab963)

[pruegg]

This is a tuff one CD34 will stain all precursor cells not just endothelial 
cells for vessels, there is an excellent CD31 for ms tissue from Dionova it is 
rat anti ms but not sure of what you could use for cd31 for rat and rabbits, 
but I know cd34 is not the way to go.

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 
rueggihcconsultin...@outlook.com



 

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: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of tahseen
Sent: Wednesday, November 06, 2013 5:34 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] CD 34 antibody abcam (ab963)

Hi All,
We used CD 34 antibody abcam (ab963)with detection kit enVision to stain our 
samples from rats and rabbits. We need to stain vessels only but macrophages, 
fibroblast, collagen all are stained in other hand known +ve control (Tonsil) 
is perform good.  Please help us that what should we do for IHC staining of our 
samples.
Best regards
Muhammad Tahseen
Sr.Supervisor Histology
SKMCH&RC
Lahore


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RE: [Histonet] FDA Disclaimer

[pruegg]

I would think so, 

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 
rueggihcconsultin...@outlook.com



 

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any of its contents, by any person other than the intended recipient may
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you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Hannen,
Valerie
Sent: Tuesday, November 05, 2013 3:48 AM
To: Histonet Post (histonet@lists.utsouthwestern.edu)
Subject: [Histonet] FDA Disclaimer

Good Morning...

I have a question about the FDA disclaimer for Immuno's.  If we are not
doing the staining of Immuno's in our lab, but our Pathologists are
interpreting those that are stained at our reference lab, are we still
required to put the FDA disclaimer on our Path reports for those antibodies
that require the disclaimer?

Thanks so much!!


Valerie

Valerie A. Hannen, MLT(ASCP),HTL,SU(FL)
Histology Section Chief
Parrish Medical Center
951 N. Washington Ave.
Titusville, Florida 32976
Phone:(321) 268-6333 ext. 7506
Fax: (321) 268-6149
valerie.han...@parrishmed.com


=
"This email is intended solely for the use of the individual to whom it is
addressed and may contain information that is privileged, confidential or
otherwise exempt from disclosure under applicable law. If the reader of this
email is not the intended recipient or the employee or agent responsible for
delivering the message to the intended recipient, you are hereby notified
that any dissemination, distribution, or copying of this communication is
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RE: [Histonet] RE: MOHS IPs

[pruegg]

   I  agree with Barbara try fixing in cold acetone/ethanol mix even = if
   just  for  a  couple  of minutes then go directly to buffer do not dry
   after=  fix,  I  would dilute the protease 1:4 with buffer (PBS or TBS
   what  ever  u  u= se) and do it for a short time.  Enzyme digestion on
   frozen  sections  i=  s  tricky  but  fixing a little and diluting the
   enzyme  should  help  prevent  ov=  er  digestion.   How  long  are  u
   incubating  the  primary?   Are  u usin= g an enhanced labeled polymer
   detection?  HRP/Dab or AEC or Alk.P/fast= red?

   


 


   
 Original Message 
   
Subject: [Histonet] RE: MOHS = IPs
   
From: Barbara Tibbs <[1]barbara.ti...@accuratediagnosticlabs.com>
   
D= ate: Wed, October 02, 2013 5:54 am
   
To: "Bauer, Karen L." <[2]bauer.ka...@mayo.edu>,
   
"'histonet@lists.utsouthwest= ern.edu'"
   
<[3]histonet@lists.utsouthwestern.edu>
   

   
Hello Karen,
   = 

   
Back  in the old days of doing IHC manually on mostly fresh, froze   n  tissue  
I would immediately place the slide with the frozen section
   into  a=  coplin  jar  of  acetone  to  fix it. Keep the coplin jar of
   acetone  in  the  cr=  yostat.  Leave  it  in  the acetone for only 10
   minutes.  Try diluting the Pr= otease 2 - 1:1 to make it gentler so it
   doesn't  "eat"  the  tissue.  This  te= chnique worked well when I was
   doing ER/PR on frozen breast tumors. Not su= re it will work with skin
   but it's worth a try.
   

   
Barbara S. Tib= bs
   
Histology Supervisor
   
Accurate Diagnostic Labs
   
South Pl= ainfield, NJ
   
[4]barbara.ti...@accuratediagnosticlabs.com
   
732-839-3374
   
C= ell: 610-809-6508
   

   

   
_= ___
   
From:  [5]histonet-boun...@lists.utsouthwestern.edu
   <[6]histonet-bounces@lists.utsouthwestern=  .edu>  on behalf of Bauer,
   Karen L. <[7]bauer.ka...@mayo.edu>
   
Sent: Tuesday, October 01, 20= 13 5:22 PM
   
To: '[8]= histonet@lists.utsouthwestern.edu'
   
Subject: [Histonet] MOHS IPs

   
Hi all,
   

   
We  are  in the process of validating some a= ntibodies in our MOHS
   lab.  The Melan A (Mart 1) antibody is working well, = but it could be
   darker.  We will be increasing the Ab incubation time to se= e if that
   will help.
   

   
As  for  the Pan Keratin, we cannot get it = to work at all. We use
   Protease  2  on  our Ultra stainer for FFPE tissues in= Histology, but
   this  stain in MOHS is placed on fresh, unfixed tissue... by= a manual
   drop  method.  Any  time we've tried to use an enzyme retrieval, th= e
   tissue looks eaten up... even if we incubate if for a minute.
   

   =  
Since  the tissue is not fixed, I figured that no retrieval step
   would  be= needed, but the Pan Keratin refuses to work with or without
   retrieval.
   = 

   
For  those of you in MOHS labs that are using a manual staining me   thod  for  
Melan A and Pan Keratin, would you be willing to share your
   protoc= ol with us?
   

   
Thanks so much!!
   

   
Karen
   

   
K= aren L. Bauer HTL/HT (ASCP) | Histology Supervisor | Pathology |
   MOHS   Lab   S=  upervisor  |  Dermatology  |  Phone:  715-838-3205  |
   [9]bauer.ka...@mayo.edu<[10]mailto:bauer.ka...@mayo.edu> | Mayo Clinic
   Health  System  |  122=  1  Whipple  Street  |  Eau Claire, WI 54702 |
   [11]mayoclinichealthsystem.org
   

   = 

   
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References

   1. 3D"mailto:barbara.tibbs@accurated   2. ="mailto:bauer.ka...@mayo.edu";
   3. 3D"mailto:histonet@lists.utsouthwestern.edu   4. 
3D"mailto:barbara.tibbs@accuratediagnosticlabs.c   5. 
3D"mailto:histonet-boun...@lists.utsouthwestern.edu   6. 3D"mailto:histo   7. 
3D"mailto:Bauer.Karen   8. 3D"mailto:histonet@lists.utsouthwestern.edu";
   9. 3D"mailto:bauer.kar  10. 3D"mailto:bauer.karen@mayo  11. 
3D"http://mayoclinichealt=/
  12. 3D"http:/  13. 3D"mailto:Histonet@lists.utsouthwestern.edu";
  14. 3D"http://lists.utsouthweste=/
  15. 3D"mailto:Histonet@lists.u  16. 
file://localhost/tmp/3D"h___
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RE: [Histonet] RE: Submitting Questions

[pruegg]

   do u see this question posted?

   


 


   
  Original Message <= br>
Subject: [Histonet] RE:
   Submitting Questions
   
From: Martha Ward-= Pathology <[1]mw...@wakehealth.edu<= /a>>
   
Date: Tue, September 24, 2013 12:54 pm
   
To:  "Nails,  Felt=  on" <[2]flnails@texaschildren= s.org>, 'Joanne
   Clark'
   
<[3]= jcl...@pcnm.com>, "[4]histonet@lists.utsouthwestern.edu"
   
<[5]histonet@lists.utsouthwestern.edu>
   

   
I have had the same problem over the past few months...
   
&n= bsp;
   
Martha Ward, MT (ASCP) QIHC
   
Manager
   

   
Molecular= Diagnostics Lab
   
Medical Center Boulevard  \  Winston-Salem= , NC 27157
   
p 336.716.2109  \  f 336.716.5890
   
<= a href="mailto:mw...@wakehealth.edu";>mw...@wakehealth.edu
   = 

   

   

   

   

   
-Original Message= -
   
From: [6]=histonet-boun...@lists.utsouthwestern.edu
   [[7]mailto:histonet-bounces@lists.utsouthweste=  rn.edu]  On Behalf Of
   Nails, Felton
   
Sent: Tuesday, September 24, 2= 013 2:58 PM
   
To: 'Joanne Clark'; [8]histonet@lists.utsouthwestern.edu
   
Subject: [Histo= net] RE: Submitting Questions
   

   
I am in the same situation??? 

   
-Original Message-
   
From:  [9]histonet-bounces@lists.utsouthwestern.=  edu  [[10]mailt   
o:histonet-boun...@lists.utsouthwestern.edu]   On   Behalf  Of  Joanne
   Clark<= br>
Sent: Tuesday, September 24, 2013 12:54 PM
   
To:  [11]histonet@lists.utsouthwestern.edu<= br>
Subject:
   [Histonet] Submitting Questions
   

   
Can  anyone  out  =  in  histoland  explain  to me why my submitted
   questions  are  not  making  it  thr=  ough to histonet? I submitted a
   question  on  the  19th  of  September, and I s= till have not seen it
   appear  in  the group emails I get daily. I sent the q= uestion to the
   correct  address  (I  checked  it  to  be  sure).  When I respond t= o
   someones question, I can see my responses, I just can't seem to get an
   or=  iginal  question  submitted.  Can  anyone tell me what I am doing
   wrong?
   = 

   

   

   

   
Disclaimer:  This electronic message may cont= ain information that
   is  proprietary, confidential, or legally privileged or= protected. It
   is intended only for the use of the individual(s) and entity= named in
   the  message.  If you are not an intended recipient of this message= ,
   please notify the sender immediately and delete the material from your
   co= mputer. Do not deliver, distribute or copy this message and do not
   disclose=  its  contents  or  take  any  action  in  reliance  on  the
   information it contains= .
   

   
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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  int= ended recipient, you are hereby notified
   that  any  review,  dissemination, o= r copying of this e-mail and its
   attachments,  if  any,  or  the  information  =  contained  herein  is
   prohibited.  If  you  have  received  this  e-mail  in error= , please
   immediately  notify  the  sender  by return e-mail and delete this e-   mail 
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References

   1. 3D"mailto:mw...@wakehealth.edu";
   2. 3D"mailto:flna...@texaschildrens.org";
   3. 3D"mailto:jcl...@pcnm.com";
   4. 3D"mailto:histonet@lists.utsouthwestern.e   5. 3D"mailto:hist   6. 
3D"mailto:histonet-boun...@lists.utsouthwestern.edu";
   7. 3D"mailto:histonet-b   8. 3D"mailto:histo...@lists.utso   9. 
3D"mailto:histon  10. 3D"mailto:histonet-boun...@lists.utsouthwestern.edu";
  11. file://localhost/tmp/3D"mailt  12. 
3D"mailto:Histonet@lists.utsouthwestern.edu";
  13. 3D"http://lists.utsouth=/
  14. ="mailto:Histonet@lists.utsouthwestern.edu";
  15. 3D"http://lists.utsouthwestern.edu/mailman/listinfo/  16. 
3D"mailto:Histonet@lists.utsouthwestern.edu  17. 3D"http://lists.utsou=/
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[Histonet] email chg for Rich Cartun

[pruegg]

Can someone help Dr Cartun make sure he is registered with Histonet?  

 

He sent me this message:

 

"Our e-mail addresses recently changed here at Hartford Hospital.  I sent an
e-mail to Histonet with "Subscribe" in the "Subject" line, but I don't know
if it was accepted.  His new email address is richard.car...@hhchealth.org 

 

 

 

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 

 

 

 

 

 


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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[Histonet] control tissues

[pruegg]

Does anyone know where I can get FFPE spleen tissue from rabbit, hamster,
and guinea pig?

 

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 

  rueggihcconsultin...@outlook.com

 

 

 

 


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] EDTA decal

[pruegg]

Good idea.  I have to soak the edta decaled tissues in an alkaline solution
inorder to restore enzyme histochemical staining for TRAP, might be the same
issue for some IHC.

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 
rueggihcconsultin...@outlook.com



 

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: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Teri Johnson
Sent: Friday, August 16, 2013 11:54 AM
To: 'cfors...@umn.edu'
Cc: histonet@lists.utsouthwestern.edu
Subject: [Histonet] EDTA decal

Hi Colleen,

I would say it's unusual, but not completely impossible that EDTA has
interfered with your IHC.  We had that problem with demonstrating
B-galactosidase in mouse bones. If we decalcified it in EDTA after whole
mount staining in X-gal, the blue staining was removed. But if we
decalcified it in formic acid, the stain was retained.

I think a few years ago, Biogenex had a room temperature antigen retrieval
solution for acid decalcified bone (not the same as your situation, I know).
But I think it was largely an alkaline solution you let the slides sit for
maybe 30 minutes prior to staining. Might be worthwhile to try a different
retrieval method for these and see what happens.

Otherwise, are you sure you are using a clone that reacts in mouse tissue?
We use Rabbit monoclonal SP6.

Teri Johnson
Manager, Histology
GNF - San Diego, CA
858-332-4752

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RE: [Histonet] IHC after EDTA decalcifying

[pruegg]

Colleen,

The antigen expression you are getting in the human tonsil may not be
expressed in the mouse femur, plus u have to make sure the ab u are using
cross reacts with mouse tissue, if it is anti human it may or may not react
in mouse, the ab data sheet should indicate species reactivity.  The edta
should not be the problem here.

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 
rueggihcconsultin...@outlook.com



 

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
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you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Colleen
Forster
Sent: Thursday, August 15, 2013 5:59 PM
To: Histonet
Subject: [Histonet] IHC after EDTA decalcifying

Hello fellow histonetters,

Can someone who has worked with decalcifyied bone tell me if EDTA interferes
with IHC staining? I was under the impression it did not but cannot get
staining in mouse femurs that I have decaled in EDTA. I have used both the
steamer and the decloaker for retrieval and in the human tonsil the stain is
great.

Any suggestions..

Thanks in advance!

Colleen Forster
U of MN

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[Histonet] processing eyes

[pruegg]

Can someone direct me to a protocol for processing eyes which have been
injected with Davidson's fixative and are now in 10% NBF.  We had an eye
expert at the U for years I was going to send my techs to but apparently she
has finally retired because we cannot get a hold of her.  Where are you Mary
Jo?


We need insight on how to process non-human primate eyes? They have been
injected with Davidson's fixative and are now in 10%NBF. We need a specific
processing schedule, embedding and sectioning advice.

 

Thank you,

 

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 

  rueggihcconsultin...@outlook.com

 

 

 

 


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
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you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.

 

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[Histonet] markers in specific species

[pruegg]

Can anyone recommend a good antibody made in rat that works in mouse tissue
that is a nuclear marker and another one that is a membrane marker?

 

Can anyone recommend a good antibody made in goat that works in mouse and
rat tissue that is a nuclear marker and another one that is a membrane
marker?

 

 

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 

  rueggihcconsultin...@outlook.com

 

 

 

 


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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[Histonet] OMICS

[pruegg]

Beware of invitations to speak at an OMICS conference, check out this link
for details

http://scholarlyoa.com/2013/01/25/omics-predatory-meetings/

 

 

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 

  rueggihcconsultin...@outlook.com

 

 

 

 


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('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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[Histonet] Mallory triple stain

[pruegg]

Does anyone know what this is about?  I think these people just want a
trichrome stain and we are already doing a Masson's TC for them using
aniline blue.  I think they are just asking for a modified version of TC and
want to make sure we use aniline blue for the collagen stain???  Is Mallory
Triple stain something other than a modified trichrome stain?

 

 

"What we were asking for was Mallory Triple stain, which appears to be the
synonymous with Mallory Aniline Blue, and Mallory Trichrome. Is that
correct, that those three names are for the same staining technique? The
main components of the stain that we were looking for were: aniline blue,
orange G, and acid fuchsin. As far as I can tell all three techniques have
those three components."

 

 

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting

40864 E. Arkansas Ave

Bennett, CO 80102

H 303-644-4538

C 720-281-5406

  rueggihcconsultin...@outlook.com

 

 

 


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('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
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any of its contents, by any person other than the intended recipient may
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you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.

 

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[Histonet] zinc salt fixed mouse tissue PE stained for CD4/cd8

[pruegg]

Has anyone tried this:  zinc salt fixed mouse tissue PE stained for CD4/cd8
I found some papers that look impressive.

 

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting

40864 E. Arkansas Ave

Bennett, CO 80102

H 303-644-4538

C 720-281-5406

  rueggihcconsultin...@outlook.com

 

 

 


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('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
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any of its contents, by any person other than the intended recipient may
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you are NOT the intended recipient please contact the sender and dispose of
this e-mail as soon as possible.

 

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[Histonet] cd8 on mouse tissue

[pruegg]

Is this still being done on frozen sections not aldehyde fixed or has
someone figured out an antibody or method of doing it on FFPE mouse tissue
yet???

 

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting

40864 E. Arkansas Ave

Bennett, CO 80102

H 303-644-4538

C 720-281-5406

  rueggihcconsultin...@outlook.com

 

 

 


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('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] RE: Antigen retrieval

[pruegg]

   The  most  important  part= about working up a new antibody is to have
   known  positive  tissue  to use as= a control, if you do not have that
   everything  you  do is a shot in the dark= , you will not know if your
   protocol  is  not  working  or  the  control  you  are  = using is not
   expressing that antigen.  Each antibody has it's own requ= irement for
   pretreatment or not.  We do what Liz does and sometimes ad= d in a ph8
   buffer  when  nothing  else works.  Just to save on slides to = run we
   start  with no ar and ph6 and if we do not get good results, go to ed   ta  
ph9,  then PK, then ph8, if all these fail, we revert to overnight
   incuba=  tion  of  the  primary  ab  after  the most agressive high ph
   buffers and/or enzy= me digestion.  If that fails we give up.  This is
   all  of  course  a=  fter the best antibody titer has been determined.
   Some of my experien= ce points to antibodies that are expressed in the
   nuclei  may  prefer  high  ph=  AR,  this  is  just  a  trend  I  have
   noticed, definitely not something I h= ave proven.

   Regards,<= /font>

   Patsy


 


   
 Original Message 
   
Subject: [Histonet] RE: Antigen retrieval
   
From: Elizabeth Chlipal= a <[1]l...@premierlab.com>
   = 
Date: Wed, June 13, 2012 7:32 am
   
To: 'Mike Tighe' <[2]mti...@trudeauinstitute.org>,
   =
"[3]histo...@lists.uts=   outhwestern.edu   ([4]h   
isto...@lists.utsouthwestern.edu)"
   
<[5]histonet@lists.utsouthwestern.edu>
   
   
Mike
   

   
Retrieval  methods  are  also  based  upon  the  antibody= . During
   protocol  development we try several different retrieval methods -= no
   retrieval,  enzymatic  (proteinase  K), pH6 and pH9, we then determine
   the=  best method and go from there. One retrieval method may not work
   for all = antibodies.
   

   
Liz
   

   
Elizabeth A. Chlipala, BS, HTL(AS= CP)QIHC
   
Manager
   
Premier Laboratory, LLC
   
PO Box 18592
   = 
Boulder, CO 80308-1592
   
(303) 682-3949 office
   
(303) 682-9060 = fax
   
(303) 881-0763 cell
   
[6]w= ww.premierlab.com
   

   
Ship to address:
   

   
1567 Skywa= y Drive, Unit E
   
Longmont, CO 80504
   

   
-Original Message= -
   
From:  [7]histonet-boun...@lists.utsouthwestern.edu
   [[8]mailto:histonet-bounces@lists.utsouthw=  estern.edu]  On Behalf Of
   Mike Tighe
   
Sent: Wednesday, June 13, 201= 2 8:27 AM
   
To:  [9]hi= sto...@lists.utsouthwestern.edu
   ([10]histonet@lists.utsouthwestern.edu)
   
Subject: [Histo= net] Antigen retrieval
   

   
Does  anyone  have  a  favorite antigen ret= rieval method for FFPE
   mouse  tissues that they would be willing to share? I= have been using
   citrate  buffer  Ph6.0  with poor to moderate results. Thanks= for any
   help!
   

   

   

   
Mike
   
__= _
   
Histonet mailing list
   
[11]Histonet@lists.utsouthwestern.edu   
[12]http://lists.utsouthwestern.edu/mailman/listinfo/histonet
   

   = 
___
   
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[13]Histonet@lis= ts.utsouthwestern.edu
   
[14]http://lists.utsouthwestern.edu/mailman/listinfo= /histonet
   



 


References

   1. 3D"mailto:l...@premierlab.com";
   2. file://localhost/tmp/3D"m   3. 
3D"mailto:histonet@lists.utsouthwestern.edu";
   4. 3D"mailto:histonet@lists.utsouthwestern.edu";
   5. 3D"mailto:histonet   6. 3D"http://www.premierlab.com"/
   7. 3D"mailto:histonet-bounces@lists.utsouthwestern.e   8. 3D"mailto:histon   
9. 3D"mailto:histonet@lists.utsouthwestern.edu";
  10. 3D"mailto:histonet@lists.utsou  11. file://localhost/tmp/3D"mail  12. 
3D"http://lists.utsouthwestern.edu/mailman/listinfo/histonet  13. 
3D"mailto:Histonet@lists.utsouthwestern.edu";
  14. 
3D"http://lists.utsouthwestern.edu/___
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RE: [Histonet] Unregistered techs

[pruegg]

   There is nothing volunteer about being ASCP certified as an HT or= HTL
   where  I have worked for the last 35 years, all those employed as HT's
   =  at  the  U of Colorado must be ASCP certified and I believe this is
   the case = for most other places doing hospital based Histology, work,
   right

   


 


   
--= -- Original Message 
   
Subject: Re: [Histonet] Unregistered tec= hs
   
From: Dorothy Ragland-Glass <[1]techman...@yahoo.com>
   
Date: Mon, May 28, 2012 10:14 pm
To: David Kemler <[2]histotalk@   yahoo.com>, Fellow HistoNetters
   
<[3]Histonet@Lists.UTSouthwestern.edu>
   
   
Dorothy R. Glass, BS,HTL(ASCP),IHC
   
  You  are  so right about = the good old days when you would prove
   who  you  were  and  sit  for  a paper not= computer exam at a medical
   school  close  to  you.  I sat for the HTL in 1988.= I was so proud to
   call  myself  a  Histologist  after  being  trained  at  a Schoo= l of
   Histotechnology  and  being ASCP certified. NOW you have people on the
   j= ob trained just referring to themselves casually as histotects. Not
   histo t= rainees. Unfortunate for us older techs, it is very offensive
   considering w= hat we went through in establishing a career not just a
   job. Some reference= labs is focusing on hiring aids to later, after a
   few  months  train  them  an= d refer to them as histotechs. I hate it
   when the term is used so freely.
Sinserely, frustrated HTL
   

   

   
David Kemler <[4]histot...@yahoo.com> wrote:
   

>Good  stuff. In the "old" days, 36 years ago for me, taking th   e  HT(ASCP) 
>exam it was said that you were "registered" by the ASC= P,
   because  the  designation HT is given by the Board of Registry of the   
ASCP. Many of the NEW folks use the word "certified". After a total o   f 39  
years  (3  years was training before you were eligible) I= still
   only  use registered by the ASCP / Licensed by the State of Florida&n   
bsp;and call myself a "histologist".
   
>
   
>In  those  d=  ays ( long before the Internet), you took your ASCP
   exam  (HT's,  MT's,  CT's,= BT's, MLT's) at specially selected medical
   schools  across  the  US. You= chose the one giving the exam which was
   closest  to  where  you  lived.  If you=  needed to drive 100 miles or
   further to  get to the examinin= g college on March 15 OR August 15th,
   (the only dates it was given)&nb= sp;that's what you did. Getting into
   the exam auditorium before y= ou were allowed to "sit" (that's what it
   was  called) for the exam, was= a challenge. You had to prove that you
   were  who  you  said  you were or you&n= bsp;were not getting in. Once
   those  guarded  doors  were closed - t= hey were CLOSED! I saw several
   folks  crying  outside the  auditorium  tha= t day n 1975. Chances for
   cheating were eliminated at every turn. Unfortuna= tely, not so today.
   So  you can  see  why  for  us older  techs, i= f you were HT(ASCP) it
   really meant something. Unfortunately, not so t= oday.
   
>
   
>Yours,
   
>Dave
   
>Hist= onetters,
   
>
   
>I see this subject tends to illicit strong s= entiments from
   
>professionals  who  are  impacted  or have an impact o= n HT/HTL's
   (sort of
   
>everyone on the net)?.
   
>
   
>I=  am  still  in  school,  but  I  want  to  fully understand how
   training,
   
>=  ;certification,  and registration work for HT/HTL’s.  I
   realize= that
   
>ASCP  certification  is  voluntary, and that some States requi= re
   some
   
>sort  of  license or certification, but I’ve never = heard
   of a “Registry”
   
>for HT/HTL’s.
   = 
>
   
>-The  way  I understand through what I’ve been taug= ht at
   school is that
   
>Histology is the study of tissue, And that..= .
   
>
   
>-To  study  tissue  there  is  another  science  that  prepa=  res
   specimens so
   
>they can be studied. And that...
   
>
   =  
>-There  is  a  final sequence “Quality Control”
   that ve= rifies the science
   
>that  prpares  specimens  is  properly  done so the= tissue can be
   studied.
   
>And that…
   
>
   
>In=  order  for  this all to happen successfully and consistently,
   the
   
>=  HT/HTL's  make  sure that during the whole preparation process,
   safety
   
= >is observed, proper adherence to federal and state regulations
   
&g=  t;maintained, plus train other technicians to do the same, and
   much
   
&= gt;more.
   
>
   
>If  I  understood  it  all  correctly  I  can��= �t help but
   wonder:
   
>
   
>If HT/HTL's do all of this cruci= al preparation work to make sure
   
>specimens  are acceptable for pre= cise microscopic identification
   of
   
>cells, tissue type, diagnosis = of disease, and other needs:
   
>
   
>"Why wouldn't we want to = have some method that can gage a set of
   basic
   
>skills to indicate = a level of competency that HT/HTL's should
   
>initially  have,  in  ord=  er to enter the field of work that can
   effect so
   
>many people eith= er directly or indire

RE: [Histonet] Unregistered techs

[pruegg]

   Well said Will.

   




   


 


    Original Message 
   Subject: Re: [Histonet] Unregistered techs
   From: William Chappell <<= a
   href="mailto:cha...@yahoo.com";>cha...@yahoo.com>
   Date: Thu, M= ay 24, 2012 4:02 pm
   To: Davide Costanzo <[1]pathloc...@gmail.com>
   Cc: histonet <[2]histonet@lists.utsouthwestern.edu>
   I  have  respected  Jay's  input  in  the  past, but I too must say s   
omething.
   Without realizing it, and by stating his opinion in a horr= ibly crass
   way,  Jay  has touched upon an important truism. There are two typ= es
   of  histologists, those that have a job that pays the bills, and those
   wh=  o have a career in which they thrive. Neither are better than the
   other,  bo=  th  are  needed. I suspect, however, that the majority of
   Histonetters  --  esp=  ecially  avid  contributors  are in the latter
   group. I know I am.
   Hist=  otechs  who  approach  histology as a job, go into work, embed,
   cut,  stain  and=  go home. they are excellent techs, but are just not
   committed  to  expanding=  the  field  or doing more than is needed to
   provide  the  pathologist  with a p= erfect slide. Jay refers to these
   people  as  no better than trained monkeys.= That is a horrible insult
   with a small (very small) grain of truth. One da= y those histologists
   will  be  replaced  by  a  mechanical/robotic  process. The = march of
   progress is unstoppable.
   The  career  histologist  has  a  much  = longer life span however. We
   analyze  and  troubleshoot  problems.  We  understa= nd or endeavor to
   learn  the organic chemistry of stains. We know EXACTLY ho= w a Rabbit
   Monoclonal  antibody  is  made.  We  know more about the practice of
histology  than  ANY pathologist. We invent and develop antibodies and
   specia=  l  stains.  And  we conceptualize and perfect the instruments
   that will replac= e the first group in the future.
   Jay,  that  is  why  so  many  are offend= ed. We don't do this simply
   because  it  is  a good paycheck. We are histologi= sts because we are
   professionals  who  choose  this  career.  You may be going t= o a job
   cutting  slides  (which is great and necessary), but we are enjoyingour 
life.
   Will Chappell, HTL (ASCP), QIHC, MBA
   and histologist by= choice, not accident
   On May 24, 2012, at 6:48 PM, Davide Costan= zo wrote:
   >  I'm  sorry - I cannot let this rest. The comment: "we = are just as
   much
   > needed as pathologists, blah, blah,
   >  blah..= ." is so upsetting I cannot sit back and listen to that
   without
   >= saying something!
   >
   >  Everyone,  regardless  of their lot in li= fe, is a very worthwhile
   part of the
   >  whole.  Let  me  ask  you a questi= on, since you highly undervalue
   humans that
   > are not MD's - let's sa= y that you are a patient at Hospital X, and
   you go
   >  in  to have your = toenail removed. Who plays a more important role
   in your
   >  survival  -=  the Podiatrist or the hospital janitor? I would argue
   that the
   >  jan=  itor is more crucial in this instance, for if he/she fails to
   clean up
   &=  gt;  the MRSA from the last patient you could conceivably die. The
   doctor so= lved
   > your fungal problem, but the janitor prevented you from gettin= g a
   >  potentially  life-threatening  infection.  Think before you speaklike 
that -
   >  everyone  involved in your care is critical - and, yes, = sometimes
   the doctor
   > is not the most important person when it comes= to keeping you alive
   and
   > well!
   >
   >
   >
   >>
   >OnThu,May   24,   2012   at   2:01   PM,   Jay   Lundgren
   <[3]jaylundg...@gmail.com> wrote:
   &= gt;
   >> Scott Lyons [4]slnj07@yah= oo.com
   >>
   >>  Give  me  a  break,  HTs and HTLs do not ma= ke diagnoses or treat
   patients. I
   >>  am  a  registered  HT  and  a Flor= ida licensed HTL with 19 years
   experience,
   >>  I've  done  it  all  in = the lab. I believe the certification and
   licensure of
   >>  techs  is  = a scam to bleed more money from people. Honestly, you
   can train a
   >&g=  t;  monkey to do our job. And I don't want to hear from everyone
   saying it's= an
   >> art form, we are just as much needed as pathologists, blah,= blah,
   >>  blah...  I  work where they are hiring people from a m= asters
   degree
   >>  program  for  histology  with  certification, THEY KN= OW NOTHING.
   Experience it
   >> where it's at, whether certified or n= ot, get off your high horse.
   >>
   >>
   >>
   >= >
   >>
   >>
   >>
   >>
   >>
   &= gt;>
   >>
   >>
   >>
   >>
   >> >>
   >>
   >>
   >>
   >>
   >>   >>
   >>> [5]Histonet@lists.utsouthwestern.edu
   >>> [6]http://lists.utso= uthwestern.edu/mailman/listinfo/histonet
   >> __= _
   >> Histonet mailing list
   >>= [7]Histonet@lists.utsout= hwestern.edu
   >> [8]http://lists.utsouthwestern.edu/mailman/listinfo/hi= stonet

RE: [Histonet] RE: processing program for biopsies

[pruegg]

   i  agree  with Liz,  the  fat  in  the  skin requires a little= longer
   processing  than  other soft tissues especially human, animals d= on't
   have  as  much  fat  and should be processed on a shorter schedule to   keep 
them from becoming too hard and dehydrated.

   


   &= nbsp;

   


 


   = Original Message 
   Subject: [Histonet] RE: processing program fo= r biopsies
   From: Elizabeth Chlipala <[1]l...@premierlab.com>
   Date:   Fri,   November   04,   2011   10:37   amTo:   'Carol  Bryant'
   <[2]cbrya@lexclin= .com>,
   "[3]hist= o...@lists.utsouthwestern.edu"
   <[4]histonet@lists.utsouthwestern.edu>
   In  my per= sonal opinion I think the processing cycle is too short. I
   know  that skin b= iopsies are tiny but we process all skin samples on
   longer  processing  cycle=  s.  Granted  I'm in research but we use 45
   minutes  to  1 hour per station for = all of our skin samples and they
   cut just fine.
   Liz
   Elizabeth= A. Chlipala, BS, HTL(ASCP)QIHC
   Manager
   Premier Laboratory, LLC
   PO= Box 18592
   Boulder, CO 80308-1592
   (303) 682-3949 office
   (303) 682-= 9060 fax
   (303) 881-0763 cell
   [5]ww= w.premierlab.com
   Ship to address:
   1567 Skyway Drive, Unit= E
   Longmont, CO 80504
   -Original Message-
   From:  [6]histonet-bounces@list=  s.utsouthwestern.edu
   [[7]mailto:histonet-boun...@lists.utsouthwestern.edu]  On  Behalf=  Of
   Carol Bryant
   Sent: Friday, November 04, 2011 10:51 AM
   To: [8]histonet@lists.utsouthwestern= .edu
   Subject: [Histonet] processing program for biopsies
   Importan= ce: High
   Would  you  experienced  histotechs mind to look at this progr= am and
   see  if it would cause skin biopsies to be poorly processed? I thinktoo 
much time in alcohols.
   The pathologists say they look bad.
   Thanks= in advance for your help!
   Station Solution Time Temp P/V Mix
   ---=  -- - - - --
   1 Formalin 0:20 OFF= P/V Slow
   2 Formalin 0:20 OFF P/V Slow
   3 70% Alcohol 0:15 OFF P/V Slo= w
   4 80% Alcohol 0:15 OFF P/V Slow
   5 95% Alcohol 0:15 OFF P/V Slow
   = 6 95% Alcohol 0:25 OFF P/V Slow
   7 100% Alcohol 0:25 OFF P/V Slow
   8 10= 0% Alcohol 0:25 OFF P/V Slow
   9 Xylene 0:35 OFF P/V Slow
   10 Xylene 0:3= 5 OFF P/V Slow
   11 Paraffin 0:15 63C. P/V Slow
   12 Paraffin 0:15 63C. P= /V Slow
   13 Paraffin 0:15 63C. P/V Slow
   14 Paraffin 0:15 63C. P/V Slow   Carol Bryant, CT (ASCP)
   Cytology/Histology Manager
   Lexing= ton Clinic
   Phone (859) 258-4082
   Fax (859) 258-4081
   [9]cb...@lexclin.com
   NOTICE OF CONFID= ENTIALITY
   This  message,  including  any attachments, is intended only = for the
   sole use of the addressee and may contain confidential or privilege= d
   information  that is protected by the State of Kentucky and/or Federal
   reg=  ulations.  If  you  are not the intended recipient, do not read,
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   have  received  this  me=  ssage  in  error,  please  call  the sender
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References

   1. 3D"mailto:liz@premierlab   2. 3D"mailto:cb...@lexclin.com";
   3. 3D"mailto:histonet@lists.utsouthwestern.edu";
   4. 3D"mailto:histo...@lists.uts   5. 3D"http://www.premierlab.com"/
   6. ="mailto:histonet-boun...@lists.utsouthwestern.edu";
   7. 3D"mailto:histonet-bounces@lists.utsouthw   8. 
="mailto:histonet@lists.utsouthwestern.edu";
   9. file://localhost/tmp/3D"mail  10. 
3D"mailto:Histonet@lists.utsouthwestern.edu";
  11. 3D"http://lists.utsouthwestern.edu/mail  12. 
3D"mailto:Histonet@lists.utsouthwestern.edu";
  13. 3D"http://lists.utsouthwestern=/
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RE: [Histonet] TGF beta

[pruegg]

   i use TGFb from Peprotech

   




   



    Original Message -= ---
   Subject: [Histonet] TGF beta
   From: "Houston, Ronald" 
   Date: Sat, October 22, 2011 8:19 am
   To: "[1]histonet@lists.utsouthwe= stern.edu"
   <[2]= histonet@lists.utsouthwestern.edu>
   Having  some  trouble  gettin=  g  any  staining with AbCam's TGF beta
   antibody? Any recommendations
   <= BR>
   Thanks
   Ronnie Houston
   = - Confidentiality Notice:
   The following mail message, includ= ing any attachments, is for the
   sole use of the intended recipient(s) an= d may contain confidential
   and privileged information. The recipient is = responsible to
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ttp://lists.utsouthwestern.edu/mailman/listinfo/histonet

References

   1. 3D"mailto:histonet@lists.utsouthwestern.edu";
   2. 3D"mailto:histonet@lists.utsouthwestern.edu";
   3. file://localhost/tmp/3D"mail   4. 
3D"http://lists.utsouthwestern.edu/mailman/listinfo/histonet";
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RE: [Histonet] Immuno. confusion

[pruegg]

   I have seen this done many ways, some people have h202 at th= e end of
   their  depara  setup  if  doing  that  off line.  For the Leica Bon= d
   instrument  we were advised to do h202 block just before the DAB and i
   hav=  e  noticed  that it works best that way for their system.  I for
   years  h=  ave  always  used h202 after the primary ab as some abs are
   affected  by  it so= I just don't take the chance of not knowing which
   may  be.   There  was = one ab, cd4 I believe that for whatever reason
   needed to be h202 blocked be= fore AR and the AB, and it really didn't
   work  if  you  did not do the block b= efore everything.  Just goes to
   show that each ab has to be handled sp= ecifically.

   




   


   A  little  aside  here and techni= cally it makes no sense but we were
   having  trouble  with  the  Leica  ap/red  ki=  t preciptating and was
   advised  by  Jim Burchette not Leica to use H202 = just before the red
   substrate chromogen and that has really improved the ap= /red staining
   for  us  which  we use a lot on derm samples.  I know = endogenous alk
   phos uses levamisol as a block and not h202 but for some rea= son this
   works so I don't argue with it.

   




   


   R= egards,

   


   Patsy

   




   


 


      Original   Message   Subject:   [Histonet]  Immuno.
   confusion
   From: "Hannen, Valerie" >
   Date: Wed, August 03, 2011 8:57 am
   To: "[1]histonet@lists.utsouthwestern.edu"
   &= lt;[2]histo...@lists.utso= uthwestern.edu>
   Hi folks...
   I  am going to try to revam= p out Immuno. procedure and am looking at
   a  couple  of  spec.  sheets  from  our=  primary  vendor. I am gettin
   conflicting information...so I am turning to y= ou all for help.
   When doing immuno's that require either HIER or enz= yme digestion, do
   you  perform  the  retrieval  first or do you do the "peroxid= e" step
   first??
   One   spec.   sheets   says   retrieve   then  block  endogenous  per   
oxidase...the  other says block endogenous peroxidase then do the HIER
   or en= zyme digestion..which is correct??
   One  other question that I have is= ...what do you use to enhance your
   stain??  The  enhancing  solution  that  we  w=  ere  using  has  been
   discontinued.
   Thanks so much!!
   Valerie Han= nen,MLT(ASCP), HTL,SU(FL)
   Histotechnologist
   Parrish Medical Center
   951 N. Washington Avenue
   Titusville, Florida 32796
   (321) 268-6111 ex= t. 7506
   = **
   "This email is intended solely for the use of the individual = to
   whom it is addressed and may contain information that is
   privilege= d, confidential or otherwise exempt from disclosure
   under applicable law= . If the reader of this email is not the
   intended recipient or the emplo= yee or agent responsible for
   delivering the message to the intended reci= pient, you are
   hereby  notified that any dissemination, distribution, or<= BR>copying
   of this communication is strictly prohibited. If you
   have rec= eived this communication in error, please immediately
   delete this messag= e. Thank you"
   **= 
   ___
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   [4]http://lists.utsouthwestern.edu/mailman/listinfo/his= tonet


References

   1. 3D"mailto:his   2. 3D"mailto:histonet@lists.utsouthwestern.edu";
   3. 3D"mailto:Histonet@lists.utsouthwestern.edu";
   4. 
3D"http://lists.utsouthwestern.edu/mail___
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RE: [Histonet] Distribution of work for older generation of Histotechs

[pruegg]

   that  is  a  very interesting thread.  I am a senior histotech = but i
   own  my own Histopathology Services Business so i get to decide w= hat
   everyone does including me.  I tend to do most of the QC/QA work a= nd
   management  these days but i do not hesitate to work at the bench when
   ne=  eded.   My  younger  assistants  are  better  suited  for cutting
   hundreds  of= sections at a time than i am these days but i do it when
   i have to.



   Regards,

   Patsy



    Original Message 
   Subject: [Hi= stonet] Distribution of work for older generation of
   Histotechs
   From:= "Gomez, Milton" <[1]Milton.Gomez= @nyumc.org>
   Date: Sun, June 05, 2011 10:09 am
   To: "'[2]histo...@lists.utsouthwestern.ed= u'"
   <[3]histone= t...@lists.utsouthwestern.edu>
   Hello Histonetters,
   Is  the=  distribution  of  work  different  for  older histotechs vs.
   younger  histotechs=  in  your  labs  and  why?  Do  they  get special
   assignments or duties because of= their growing wisdom and seniority?
   Thanks in advance,
   MG
   
   
   
   --= --
   This  email  message, inclu= ding any attachments, is for the sole use
   of  the  intended  recipient(s)  and = may contain information that is
   proprietary,   confidential,   and  exempt  from  =  disclosure  under
   applicable  law.  Any  unauthorized  review,  use,  disclosure,  =  or
   distribution  is  prohibited. If you have received this email in error
   ple=  ase  notify  the  sender by return email and delete the original
   message.  Plea= se note, the recipient should check this email and any
   attachments for the = presence of viruses. The organization accepts no
   liability  for  any  damage  c= aused by any virus transmitted by this
   email.
      ===   3D== 
   
   
   
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References

   1. 3D"mailto:milton.go...@nyumc.org";
   2. file://localhost/tmp/3D   3. 3D"mailto:histonet@lists.utsouthwestern.edu";
   4. ="mailto:Histonet@lists.utsouthwestern.edu";
   5. 
3D"http://lists.utsouthwestern.edu/mailman/listinfo/his___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
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RE: [Histonet] Control Slides

[pruegg]

   I do not melt dry my cut control slides until i need to use them,= and
   i  do store them in boxes at 4c, i have recently run into this problem
   =  with  blocks,  so  now i seal all my blocks with paraffin and store
   them in th= e fridge as well.



   Patsy



    Original Message 
   Subject: RE:= [Histonet] Control Slides
   From: "Harrison, Sandra C." <[1]sandra.harris...@va.gov>
   Date: Mon,= March 14, 2011 1:40 pm
   To: "Lee & Peggy Wenk" <[2]lpw...@sbcglobal.net>, "Amador, Amanda"
   &l=t;[3]aama...@ameripath.com>,&=   lt;[4]histo...@lists.utso   
uthwestern.edu>
   We  dry  the  control  slide at the same time tha= t we dry the tissue
   being
   stained,  since  controls  are  supposed  to  be han= dled in the same
   manner as
   the test tissue. That being said, I guess if = we were truly following
   that rule, we would cut the control on the same = day we cut the test
   sample. :-)
   "Other people say that, after   they  dry  the  slide,  they  dip  the slide 
in paraffin, to cover the
   tissu= e,
   so
   that  the  air  doesn't touch the tissue during the months before= the
   slide
   is
   used in a stain."
   -Original Message-
   From: [5]histone= t-boun...@lists.utsouthwestern.edu
   [[6]mailto:histonet-bounces@lists.utsouthwestern.e=  du]  On Behalf Of
   Lee &
   Peggy Wenk
   Sent: Thursday, March 10, 2= 011 6:39 PM
   To: Amador, Amanda; [7]histonet@lists.utsouthwestern.edu
   Subject: Re: [Histone= t] Control Slides
   I've  noticed  that  our  spirochete  control  slides do= n't stain as
   intense
   starting about 3 months after they are sectioned, = and that they stop
   staining by about 6 months. (This is with a silver s= tain.)
   I've  talked  with  other  histotechs, and they say they've seen = the
   same
   phenomenon with AFB and Gram controls. (We use ours up too qui= ckly -
   we
   never  have  6-12  months  old  control slides for these, so I can= 't
   attest
   to
   this. )
   Bancroft's  book  talks  about  this  phenome=  non  with amyloid, and
   suggests
   that
   oxidation of the tissue (protein= s) due to exposure to air may be the
   cause.
   I'm  guessing  the  it  prob= ably applies to the precut microorganism
   control
   slides, too.
   We  only  cut  enough slides for 6 months, and date them. Other people
   say
   they
   put their cut control slides in a slide box with a lid after dryi= ng,
   and
   then place the box in the refrig, as cold slows down the che= mical
   change,
   and  the  lid  keeps  the moving air off the slide. Other = people say
   that,
   after
   they  dry  the  slide,  they dip the slide in par= affin, to cover the
   tissue,
   so
   that  the  air  doesn't touch the tissue= during the months before the
   slide
   is
   used in a stain. Just 3 sugge= stions to stop this problem.
   Peggy A. Wenk, HTL(ASCP)SLS
   Beaumont= Hospital
   Royal Oak, MI 48073
   ---= ---
   From: "Amador, Amanda" <[8]aama...@ameripath.com>
   Sent: Wednesday, March 09, 201= 1 10:22 PM
   To: <[9]= histonet@lists.utsouthwestern.edu>
   Subject: [Histonet] Control Sl= ides
   >  Is there guidelines for how long special stains controls a= re good
   for
   once
   >  they  are  cut?  We have spirochetes for our Stei= ner that is from
   2007
   and
   > we are having issues.
   >
   >= Amanda Amador, HT(ASCP)CM
   >  AmeriPath  |  Histology  Group Lead/Trainer= |2560 N Shadeland Ave,
   Suite
   A |
   > Indianapolis, IN 46219 | phon= e 317.275.8052 |
   > [10]aamador@= ameripath.com; |
   > [12]www.AmeriPath.com;
   >
   > ___   > Histonet mailing list
   > [13]Histonet@lists.utsouthwestern.edu
   > [14]http://lists.utso= uthwestern.edu/mailman/listinfo/histonet
   __= _
   Histonet mailing list
   [15]Histonet@lists.utsouthwestern.edu[16]htt   
p://lists.utsouthwestern.edu/mailman/listinfo/histonet
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References

   1. file://localhost/tmp/3D"ma   2. file://localhost/tmp/3D"mailto   3. 
3D"mailto:aama...@ameripath.com";
   4. 3D"mailto:histonet@lists.utsouthwestern.edu";
   5. 3D"mailto:histonet-boun...@lists.utsouthwestern.edu";
   6. 3D"mailto:histonet-bounc   7. 3D"mailto:histonet@lists.utsouth   8. 
3D"mailto:aamador@ame   9. 3D"mailto:histonet@lists.utsouthwestern.edu";
  10. 3D"mailto:aama...@ameripath.com";
  11. 3D"http://aama...@ameripath.com>"/
  12. 3D"http://www.AmeriPath.com&l=/
  13. 3D"mailto:histo...@lists.uts  14. file://localhost/tmp/3D"h  15. 
file://localhost/tmp/3D"mailto  16. 
3D"http://lists.utsouthwestern.edu/mailman/listinfo/histonet";
  17. ="mailto:Histonet@lists.utsouthwestern.edu";
  18. 
3D"ht

RE: [Histonet] antigen retrieval-pressure cooker-CD44- CD24-canine tissue

[pruegg]

   Anjan,

   I  am  more  of a fan of steamer or waterbath than pressure cooker, in
   my=  hands  at altitude the pressure cooker can be very harsh and make
   my  tissue=  s  fall off the slide or get chewed up badly.  I know the
   pressure  is  s= upposed to be controled by the cooker but in my hands
   boiling  seems to be t= aking place, violent boiling and i do not like
   it.

   are  you  sure  the  antibodies work on canine tissue?  When testingabs, 
I have four protocols I run

   1. no pretreatment

   2. HIER with lph citrate buffer

   3. HIER with hph buffer, such as edta

   4. EIER, I will start with proteinase K for 5min.

   usually  with  one of these, if the ab x reacts with canine tissue you
   w=  ould  get  some  signal,  if  it  is  weak  i  would  increase the
   pretreatment  times= for that one approach.  If i get nothing from any
   of them and I know = I am using the optomal dilution of the primary ab
   for  the  longest  incubatio=  n  time  (sometimes  i go to over night
   incubation  of primary) then I really w= onder if that ab works on the
   species I am trying to use it on.



   Cheers,

   Patsy



    Original Message 
   Subject:   [Histonet]   antigen   r=   etrieval-pressure  cooker-CD44-
   CD24-canine
   tissue
   From: anjan kumar &= lt;drvet_an...@hotmail.com>
   Date: Sun, May 24, 2009 1:48 pm
   To: tr= iple immunohistochem 
   h= ello everyone,
   I  wanted  to know the exact stepwise protocol for antigen = retrieval
   using  a pressure cooker. i tried using some protocol but there is= no
   staining.  Kindly  tell  me  the detailed protocol, like when i should
   plac=  e  the  slides  into the pressure cooker and from when i should
   count the time= .
   And  i  am  using  mouse monoclonals CD44 & CD24 labvision in can= ine
   tissue,  has  anyone  got  experience with these antibodies as citrate
   buff= er doesnt at all seems to work.
   plz send me some information on this.
   Regards,
   Dr. Anjan Kumar.K.R
   M.V.Sc Scholar
   Dept. of = Veterinary Pathology
   Madras Veterinary College
   Chennai-7
   <= BR>India
   email: drvet_an...@hotmail.com
   Phone: +91-9940475801   
___= __
   Live  Search extreme As India feels the heat of poll season, get a= ll
   the info you need on the MSN News Aggregator
   [1]http://news.in.msn.com/National/indiael   
ections2009/aggregator/default.aspx___
   _= ___
   Histonet mailing list
   histo...@lists.utsouthwestern.edu[2]http://lists.utsouthwestern.edu/ma
   ilman/listinfo/histonet
   <= /DIV>

References

   1. 3D"http://news.i=/
   2. 3D"http://lists.utsouthwestern.edu/mailman/listinfo/histonet";
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RE: [Histonet] Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does

[pruegg]

   i  have a protocol from Bryan Hewlett that is called "the fail pro= of
   Masons Trichrome"

   Patsy



    Original Message 
   Subject: RE: [Histonet] Does = anyone know how to achieve a perfect
   Masson  Trichrome  stain.  Our  counter=  stain (light green in acetic
   acid)
   works well but the intensity of the m= uscle stain is not very bright.
   Does anyone have any suggestions? We wil= l be staining liver
   From: "Lee & Peggy Wenk" 
   Date: Tue, December 30, 2008 3:20 pm
   To: "'SARAH REEVES'" ,
   
   Lots of questions, as this is one of those stains that has a lot of p   laces
   that need to be tweaked:
   NEW OR OLD? Is this a new problem = you are having, or have you always
   had
   this problem?
   MORDANT: Are= you still post-mordanting in Bouin, or have you tried to
   eliminate  this=  or  switch  over to something else to try to not use
   picric
   acid in the l= ab? What time and temperature?
   RED  DYE:  What dye(s) are you using fo= r the red dye for the muscle?
   Please
   include name and color index (CI) n= umber, if that's available in the
   UK.
   Also,  have  you  switched  recently  t=  o  a new type of red dye, or
   purchased a
   replacement when you ran out? Wh= at is the dye content percent on the
   dry
   powder?
   PMA/PTA:  Are  you=  using  Phosphotungstic acid and/or Phospomolybdic
   acid? How
   old is it?
   ONE OR TWO STEP: Just to make certain, is this a true Masson trichrom   e,
   where the red dye is applied, and then the PTA/PMA, and then finallythe
   green  dye? Or is this the traditional Gomori, where the red and gree   n 
dyes
   and the PTA/PMA are all in one solution?
   TIME  IN  RED AND G= REEN DYES: Are you trying to standardize the time
   in the
   red  dye  and  the=  time in the green dye? This usually doesn't work.
   Each
   different  type  o=  f  tissue,  and each different different person's
   tissue, will
   pick up red= dye and green dye at a different rate.
   TEMPERATURE:  Are the dyes a= t room temp or refrigerator temperature?
   That can
   have an influence.
  MACHINE  OR  HAND:  Are you hand staining, where you have control over
   the   amount of time in each solution, where you can check the progress with
   = a
   microscope,  where you have a choice of dyes? Or are you staining with
   = a kit?
   Or on a machine?
   Sorry,  need  some  more information, before= we can help you pinpoint
   the
   problem.
   Peggy A. Wenk, HTL(ASCP)SL= S
   Beaumont Hospital
   Royal Oak, MI 48073
   -Original Message-= 
   From: histonet-boun...@lists.utsouthwestern.edu
   [[1]mailto:histonet-boun...@lists.utsouthwestern.edu]  On  Behalf O= f
   SARAH REEVES
   Sent: Tuesday, December 30, 2008 10:09 AM
   To: histonet= @lists.utsouthwestern.edu
   Subject: [Histonet] Does anyone know how to ac= hieve a perfect Masson
   Trichrome  stain. Our counterstain (light green in= acetic acid) works
   well
   but  the  intensity  of  the  muscle  stain is not ve= ry bright. Does
   anyone have
   any suggestions? We will be staining liver b= io
   Does anyone know how to achieve a perfect Masson Trichrome stain.= Our
   counterstain  (light  green in acetic acid) works well but the intens   ity 
of
   the  muscle  stain  is  not very bright. Does anyone have any suggest   
ions? We
   will  be  staining liver biopsies mostly in the near future and w= ish
   to
   improve our technique.
   Thanks in advance
   Sarah
   <= BR>
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   This email and any files transmitted with it are confidential and int   ended
   solely  for  the  use of the individual or entity to whom they are ad   
dressed.
   Any  views  or opinions presented are solely those of the author = and
   do not
   necessarily  represent  those  of  East Kent Hospitals University= NHS
   Trust. If
   you  are  not  the  intended  recipient,  be  advised  that you h= ave
   received this
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   or
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RE: [Histonet] Silly Question?

[pruegg]

   i  have had this argument with the researchers at the University f= or
   30  years,  somewhere back in the day they were told that commercially
   mad= e formalin had methanol in it (it does but just a little and does
   not  hurt  = anything in my experience) and that methanol would damage
   their  tissue  for = IHC, so they think they must use paraformaldehyde
   and make it fresh themsel= ves.  Since new people make it all the time
   it  often  does  not get= made up correctly and their stress over this
   issue  is  miss  placed as they = should be using something commercial
   for  consistancy  and  paying  more  attent=  ion to adequate time for
   fixation in reg formalin.

   Another  anoying  myth  that  is difficult to combat with them is that
   "we=  should  limit the fixation time in aldehyde fixatives because it
   will  cross=  link the proteins masking them for IHC", there fore i am
   always  getting ti= ssue that has not been fixed long enough (at least
   24  hrs.  to  protect  it  fr=  om paraffin processing, because if the
   proteins  are  not  cross  linked  they  c= an be alcohol fixed and/or
   washed  away  forever),  the people in research kno= w about the cross
   linking  fo aldehydes but do not know that cross linking o= f proteins
   is  a  good  thing  and  they also do not know that we have advanced
methods HIER or EIER for unmasking the proteins, but we have no way of
   gett=  ing a protein back that has been lost in processing because the
   sample was = not adequately fixed.



   there i will get off my Friday soap box..



   Happy Holidays to all!



   Patsy



    Original Message 
   Subject: RE: [Histonet] Silly= Question?
   From: Merced Leiker 
   Date: Fri, = December 12, 2008 8:12 am
   To: "Edwards, R.E." 
   Cc: "'histo...@lists.uts= outhwestern.edu'"
   
   In  re= search lab situations particularly, one does not have the time
   or
   techn= ique for nailing down the ways of making each of the buffers,
   reagents,=  and  procedures  work  the "right" way or the most optimum
   way...a
   lot of= times it's students or postdocs just focused on getting their
   project = done and not caring how their fixative is made as long as it
   "works" to= some degree and, alas, it's up to us already over-booked
   technicians  t=  o  figure  out the best way to make the PFAand we
   usually
   don't have = a whole day (week, or year) to spend researching the
   back-and-forth arg= uments, either! ;-)
   Merced
   --On Friday, December 12, 2008 2:0= 4 PM + "Edwards, R.E."
wrote:
   &=  gt; You hit the nail on the head "That's what we always use", fear
   of
   &g= t; change is a common human condition.
   >
   > -Original Messag= e-
   > From: histonet-boun...@lists.utsouthwestern.edu
   >  [[1]mailto:histonet-boun...@lists.utsouthwestern.edu]= On Behalf Of
   Pat
   > Flannery Sent: 11 December 2008 16:59
   > To:= histonet@lists.utsouthwestern.edu
   > Subject: [Histonet] Silly Questi= on?
   >
   >  Please humor me on this if it's obvious (to everyone bu= t me): why
   do
   >  we  use  paraformaldehyde  (which  is so inconvenient to = make up)
   rather
   > than buffered formalin or just diluted formaldehyde= itself?
   >
   >  It  seems  that  around  here, some folks prefer paraf= ormaldehyde
   (either
   >  2%  or  4%)  and  others use formalin, while some o= thers stick to
   diluted
   > formaldehyde (I see all 4 on labels for spec= imens submitted for
   >  histology). Is it mostly a matter of personal p= reference or where
   you
   >  were  trained (i.e. force of habit) or is the= re a valid reason to
   use
   > each solution (basically the same chemical= once in solution, merely
   > buffered or not)? The only answer I've go= tten when I've asked is,
   > "That's what we always use."
   >
   &g= t; Thanks.
   >
   >  -Pat  Flannery  (not a "real" histologist - I just= play one in the
   lab)
   >
   >
   > _= __
   > Histonet mailing list
   > histo...@lists.uts= outhwestern.edu
   > [2]http://lists.utsouthwestern.edu/mailman/list= info/histonet
   >
   > _= __
   > Histonet mailing list
   > histo...@lists.utsouthwestern.= edu
   > [3]http://lists.utsouthwestern.edu/mailman/listinfo/histone= t
   >
   Merced M Leiker
   Research Technician II
   3= 54 BRB (pkgs) / 140 Farber Hall (letters)
   School of Medicine and Biomedi= cal Sciences
   State University of New York at Buffalo
   3435 Main St, Bu= ffalo, NY 14214
   Ph: (716) 829-6033
   Fx: (716) 829-2725
   "Without= my flaws I'm really very boring."
   - random internet blog commentator
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References

   1. 3D"http://email.secureserver.=/
   2. 3D"http://lists.utsouthwest

[Histonet] posting for Tim

[pruegg]

   Subject:
   respond directly to Tim on this at&nbs= p;[1]tmal...@cox.net



   HELP ! HISTONET ADDRESS
   From:
   "Timothy Malloy" <[2]tmal...@cox.net> ([3]Add as Preferred Sender)
   [4] 3D?
   <= /TD>
   Date: Thu, Dec 11, 2008 2:57 am
   To: <[5]pru...@ihctech.net>, <[6]tmallo...@hotm= ail.com>,
   <[7]tmal...@cox.net>

   Hi Patsy,
   I am a fan of yours on histonet. Just recently I moved my mail fr= om
   hotmail to outlook and lost histonets address. Could you supply me
   with = this request. I want to post a question . maybe you would be
   likely to answ= er this one. We work for the providence RI VA Hospital
   and we are in need o= f a procedure for GMS using a hot plate instead
   of a microwave. We have iss= ues venting the microwave because of the
   age of the building. Would you kno= w of a silver stain to test for
   pneumocystisis on a hot plate??

   Thank You Tim Malloy

   [8]tmal...@cox.net or
   [9]tmallo...@hotmail.= com

   Timothy Malloy  HT  (ASCP) A.A.S.

References

   1. 3D"http://em=/
   2. 3D"http://email.secureserver.net/addressBookQuickAdd.php?contact   3. 
3D"http://email.secureserver.net/w   4. 3D"http://email.secureserver.net/webm   
5. 3D"http://email.secureserver.net/addressBookQuickAdd.php?cont   6. 
3D"http://email.secureserver.net/addr   7. 3D"http://emai=/
   8. file://localhost/tmp/3D"m   9. 3D"mailto:tmallo...@hotmail.com";
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RE: [Histonet] de-waxing / staining station question

[pruegg]

   i  would  definately  always  use at least 2 xylenes and change the f   irst 
 often  where most of the paraffin is removed, or at least rotate
   it and= put a clean xylene at the end



    Original Message 
   Subject: [Histonet] de-waxing= / staining station question
   From: "Anna K. Schultz" <[EMAIL PROTECTED] .edu>
   Date: Wed, December 10, 2008 1:12 pm
   To: [EMAIL PROTECTED] uthwestern.edu
   Hi all,
   In an effort to reduce space usage, it has= been suggested that I
   eliminate one of two xylene steps (and lengthen = the time in the
   remaining xylene) when de-waxing paraffin sections. I d= o not see any
   protocols that only use one step of xylene. Are two stric= tly
   necessary? I would like to know the reasoning.
   Also, do gill hem= atoxylins and alcoholic eosin need to be used and
   stored under a hood? = I've read the MSDS but it wasn't entirely clear
   if
   they  should  just  be = used with caution or strictly under the hood.
   They
   are in staining buck= ets with lids (the lids are sometimes leaky).
   I thought I would seek som= e advice here.
   Thanks,
   Anna
   basic science research core tech
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   Histonet mailing list<= BR>Histonet@lists.utsouthwestern.edu
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References

   1. 3D"http://lists.utsouthwest=/
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