RE: [Histonet] Routine H & E stain,

[amitapandey]

We are using Xylene sulphur free- LR grade( from Rankem, India). I don't 
know about its synthetic nature but i will inquire this to manufacture.
Mountant DPX from Merck .

Do you have any suggestion on this combination?

Thanks for all your feed back.

Amita



From:   
To: 
Cc: 
Date:   07/07/11 07:10 PM
Subject:RE: [Histonet] Routine H & E stain,



I have always used Harris and have not experienced this problem?  I do
buy mine pre-made from VWR (cat#-95057-858).  I agree with the xylene
comment, also what mounting media are you using?  The mounting media can
fade your stain over time as well. 
Good Luck!

Sarah Goebel-Dysart, BA, HT(ASCP)
Histotechnologist
Mirna Therapeutics
2150 Woodward Street
Suite 100
Austin, Texas  78744
(512)901-0900 ext. 6912


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
amitapan...@torrentpharma.com
Sent: Wednesday, July 06, 2011 11:23 PM
To: Histonet; histonet-boun...@lists.utsouthwestern.edu; histonet
Subject: [Histonet] Routine H & E stain,

Hello Histonetters,

I am from toxicopathology lab where we perform H&E on rat tissues and 
store these slides for 10 long years.

We use self lab prepare Harris hematoxylin (Hematoxylin crystal+Alcohol+

Ammonium alum +D/W and Mercuric oxide for ripening) and 1% aqueous
eosin. 
The staining result is good (Purplish pink look stained slide) on all 
tissue, but  our observation is after 5-6 months time this get faded and

become towards pinkish type though we can observe the slide. 
We want to retain its fresh stained color. 

Please suggest me how to keep stable stain for longer period or do you 
suggest to switch to any other type of haematoxylin - prepared or 
commercially available?

Looking forward for your feed back.
Amita
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[Histonet] low profile vs high profile blades

[Liette Tougas]

Hi everyone,

Maybe because I had been using the same type all along (low profile) but I was 
just wondering what is the technical reason for having the two types of blades? 
 One cuts better than the other? Price? Longer lasting?

Thank you in advance for your feedback,

Liette Tougas, RT, B.Sc., M.Sc.
Biomedical Laboratory Technology Department
Dawson College, Montreal, Canada


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[Histonet] microtome knife holder

[Liette Tougas]

Hi again everyone,

I also wanted to ask if anyone has, or new if there was ever, a regular knife 
(not blade) holder for the Reichert Yung 2030 microtome and/or the Leica 
microtome series.

thank you again in advance,

Liette Tougas, RT, B.Sc., M.Sc.
Biomedical Laboratory Technology Department
Dawson College
514-931-8731, ext 1519
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[Histonet] RE: Stripped H&E's - HELP!

[Weems, Joyce]

Are you sure the alcohol covers the slides well? Sounds like water running from 
the top of the slide. 


Joyce Weems 
Pathology Manager 
Saint Joseph's Hospital 
5665 Peachtree Dunwoody Rd NE 
Atlanta, GA 30342 
678-843-7376 - Phone 
678-843-7831 - Fax 


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jo-Ann Bader, 
Ms.
Sent: Thursday, July 07, 2011 15:38
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Stripped H&E's - HELP!

Hi All,

Over the past year we have had a recurring problems with our H&E's.  The come 
out of our Gemini Stainer stripped but not on every run; we can go for months 
without a problem and then all of a sudden it is back again.  The stripping is 
in the Eosin, our pathologist confirms the the hemotoxylin is not affected.  
There are 3 of us cutting on 3 different microtomes.  The age on the microtomes 
range in age from 10 years to less than 1 year. 

Some of the solution we have tried are:

We numbered all our racks to see if some of our older racks may have micro 
cracks in them that may retain solution which gradually dripped onto the 
slides.  After checking for over a month there was no pattern to  racks and bad 
staining.

We tried lowering the level of the solutions to below the coated end of the 
slides.  No results

We tried changing the positions of the pots for the solutions,  thinking that 
if there was a leak from the solutions above we would notice a difference.  No 
luck.

We checked to see if the stripes only appeared near the end of our stains life. 
 Not that either

Right now I have a technician sitting in front of the stainer as it runs to see 
if she can catch any abnormalities in the way the machine is transferring the 
slides.

We rotate our alcohols every run.

We change every solution after every 450 slides.

I have put a call into the company to see if I can get to speak to a specialist 
on this machine but in the mean time I was hoping someone out there could help. 
 Has anyone seen such a problem

I, we, have run out of ideas aside from it being the machine and not being able 
to find out exactly how.


Jo-Ann Bader
Histology Co-Ordinator
Goodman Cancer Centre
McGill University 1160 Pine Ave. W - Rm 312 (3355) Montreal, QC, Canada H3G 1Y6
Tel: 514-398-8270
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RE: [Histonet] Stripped H&E's - HELP!

[Laurie Colbert]

Make sure the level of the alcohols after the eosin covers the tops of
the slides.  It's most likely water dripping down the slides, and this
cause the eosin on the sections to bleed so they looked striped.
Laurie Colbert

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jo-Ann
Bader, Ms.
Sent: Thursday, July 07, 2011 12:38 PM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Stripped H&E's - HELP!

Hi All,

Over the past year we have had a recurring problems with our H&E's.  The
come out of our Gemini Stainer stripped but not on every run; we can go
for months without a problem and then all of a sudden it is back again.
The stripping is in the Eosin, our pathologist confirms the the
hemotoxylin is not affected.  There are 3 of us cutting on 3 different
microtomes.  The age on the microtomes range in age from 10 years to
less than 1 year. 

Some of the solution we have tried are:

We numbered all our racks to see if some of our older racks may have
micro cracks in them that may retain solution which gradually dripped
onto the slides.  After checking for over a month there was no pattern
to  racks and bad staining.

We tried lowering the level of the solutions to below the coated end of
the slides.  No results

We tried changing the positions of the pots for the solutions,  thinking
that if there was a leak from the solutions above we would notice a
difference.  No luck.

We checked to see if the stripes only appeared near the end of our
stains life.  Not that either

Right now I have a technician sitting in front of the stainer as it runs
to see if she can catch any abnormalities in the way the machine is
transferring the slides.

We rotate our alcohols every run.

We change every solution after every 450 slides.

I have put a call into the company to see if I can get to speak to a
specialist on this machine but in the mean time I was hoping someone out
there could help.  Has anyone seen such a problem

I, we, have run out of ideas aside from it being the machine and not
being able to find out exactly how.


Jo-Ann Bader
Histology Co-Ordinator
Goodman Cancer Centre
McGill University 1160 Pine Ave. W - Rm 312 (3355)
Montreal, QC, Canada
H3G 1Y6
Tel: 514-398-8270
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[Histonet] RE: Tissue Cassette Printer Recommendations

[Rodriguez, Arnold]

All very good suggestions and definitely things to consider. Thank you
Jesus.

-Original Message-
From: Jesus Ellin [mailto:jel...@yumaregional.org] 
Sent: Thursday, July 07, 2011 10:05 AM
To: 'Carol Bryant'; 'Elizabeth Chlipala'; Rodriguez, Arnold;
histonet@lists.utsouthwestern.edu
Subject: RE: Tissue Cassette Printer Recommendations

I do not have Vantage, so I cannot comment on that. But you have to take
into account the environment and the scanning quality of the print.  I
would also look at space, location, downtime, workflow, even clarity of
barcode meaning Datamatirx, ect.  There are also issues with specific
cassette colors and cassette surface texture playing a part in the
scanning.

-Original Message-
From: Carol Bryant [mailto:cb...@lexclin.com]
Sent: Thursday, July 07, 2011 9:55 AM
To: 'Elizabeth Chlipala'; Jesus Ellin; 'Rodriguez, Arnold';
histonet@lists.utsouthwestern.edu
Subject: RE: Tissue Cassette Printer Recommendations

I am intersted in this info also.  I would like to know what printers
everyone are using with the Vantage system or a bar coding system. 
Thank you,
Carol 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
Elizabeth Chlipala
Sent: Thursday, July 07, 2011 12:53 PM
To: 'Jesus Ellin'; 'Rodriguez, Arnold';
histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Tissue Cassette Printer Recommendations

I second Jesus's comments especially if you are moving towards bar
coding.  You will need a cassette printer that can print a high quality
2D bar code.

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
www.premierlab.com

Ship to address:

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 Jesus
Ellin
Sent: Thursday, July 07, 2011 10:49 AM
To: 'Rodriguez, Arnold'; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Tissue Cassette Printer Recommendations

I would look at Thermo, Leica and General Data printers .

Jesus Ellin
Yuma Regional Medical Center

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
Rodriguez, Arnold
Sent: Thursday, July 07, 2011 9:38 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Tissue Cassette Printer Recommendations

Hello All,

We are interested in purchasing a new cassette printer and I would
sincerely appreciate any recommendations for this instrument. We are
particularly looking for reliability, print speed, LIS connectivity and
barcode technology.

Thank you very much.

Arnold Rodriguez, HT (ASCP)
Supervisor, Anatomic Pathology
Eisenhower Medical Center
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Re: [Histonet] pax2

[Mark Tarango]

The only problem here is that the Cell Marque antibody is currently on
backorder until at least the end of August from what I've heard.

Mark

On Thu, Jul 7, 2011 at 12:36 PM, Angela Bitting wrote:

> CC1 standard, 32 min incubation with heat disabled. I use Cellmarques
> predilute
>  and UltraView DAB detection.
>
> >>> Dorothy Glass  7/7/2011 3:28 PM >>>
>  Does anyone have a working protocol for pax2 on the Ventana Ultra or XT?
>  Can you please share on Histonet?
>
> Dorothy Glass
> Supervisor, SEPA LABS
> Brunswick,Ga.
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RE: [Histonet] RE: Tissue Cassette Printer Recommendations

[Rodriguez, Arnold]

Thank you for the prompt reply. I was also told that the Sakura and
Leica printers are the same. Speed of printing is close to the top of
our least, so thank you for that bit of information as well. 

 

Have a wonderful day.

 

Arnold Rodriguez, HT (ASCP)

Supervisor Anatomic Pathology

Eisenhower Medical Center 




From: CHRISTIE GOWAN [mailto:christiego...@msn.com] 
Sent: Thursday, July 07, 2011 10:22 AM
To: jwe...@sjha.org; Rodriguez, Arnold;
histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] RE: Tissue Cassette Printer Recommendations



 The Sakura and the Leica are the same system as they share a patent
(this is what I am told). We use the Leica IPC for our cassettes and we
have the Vantage system. We did a demo of the Thermo printer but were
not happy with the speed and the fact that we would need to change who
we order cassettes from. We did change the color of our routine
cassettes from blue to white as the white scanned better at the vantage
stations. We still use blue but only for autopsy cases. We use the
Vantage system at the printer to scan in the patient bar code, choose a
hopper with the color we want and hit print all. So far it has worked
quite well.
Christie Gowan
cgo...@uabmc.edu
 

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[Histonet] Secondary antibody for Ventana detection kits

[Véronique Barrès]

Hi,

I would like to know if you finally optimized your staining with the
anti-goat and the open secondary kit. If yes, can you tell me which antibody
and dilution you used?
I'm currently trying to do the same thing here. So far I only tried
secondary antibodies from santa-cruz (the same that we use for western blot)
without any positive result.

Thanks,

Véronique Barrès, Research Assistant

Institut du cancer de Montreal
Phone: 514-890-8000 ext. 24647
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RE: [Histonet] RE: Specimen Collection Instructions

[CHRISTIE GOWAN]

We have a website at our hospital that is accessible to all physicians and 
nursing staff for collection of all specimens. We call the website LabSource.
 

> From: trathbo...@somerset-healthcare.com
> To: mpe...@grhs.net; histonet@lists.utsouthwestern.edu
> Date: Thu, 7 Jul 2011 19:36:15 +
> CC: 
> Subject: [Histonet] RE: Specimen Collection Instructions
> 
> We have the basic specimen requirements (taken from our procedure manuals) 
> from each of the Lab areas, and place them in a binder for each of the 
> Nursing units. So whether it is a fluid or a tissue specimen, or if it 
> requires special handling or collection, it is in this manual. If something 
> arrives collected improperly, the unit manager is notified and a specimen 
> incident report is generated.
> 
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu 
> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mike Pence
> Sent: Thursday, July 07, 2011 2:55 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Specimen Collection Instructions
> 
> How has everyone handled this question GEN.40104 Instructions are distributed 
> to physicians and paramedical personnel for proper collection, handling, 
> transportation, and preparation of cytologic and tissue specimens.
> 
> I feel this question is asking me to spell out to a Dr how to collect his/her 
> specimens. To me this is like telling him/her how to practice medicine.
> 
> I hope someone would like to share what you have seen that works.
> 
> Thanks, Mike
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[Histonet] RE: Specimen Collection Instructions

[Rathborne, Toni]

We have the basic specimen requirements (taken from our procedure manuals) from 
each of the Lab areas, and place them in a binder for each of the Nursing 
units. So whether it is a fluid or a tissue specimen, or if it requires special 
handling or collection, it is in this manual. If something arrives collected 
improperly, the unit manager is notified and a specimen incident report is 
generated.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mike Pence
Sent: Thursday, July 07, 2011 2:55 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Specimen Collection Instructions

How has everyone handled this  question GEN.40104 Instructions are distributed 
to physicians and paramedical personnel for proper collection, handling, 
transportation, and preparation of cytologic and tissue specimens.
 
I feel this question is asking me to spell out to a Dr how to collect his/her 
specimens. To me this is like telling him/her how to practice medicine.
 
I hope someone would like to share what you have seen that works.
 
Thanks, Mike
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[Histonet] HT Position in Las Vegas

[Kaitlin Webster]

Histology position in Las Vegas, Nevada with national reference lab. The
position is a night shift and includes $5k sign-on bonus as well as
relocation package. Pease contact me for immediate consideration. 

 

 

Kaitlin Webster

Account Manager 

Prometheus Healthcare

Office (301) 693-9057

Cell (407) 334-4438

Fax (301) 368-2478

kait...@prometheushealthcare.com

www.prometheushealthcare.com

 

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[Histonet] Stripped H&E's - HELP!

[Jo-Ann Bader, Ms.]

Hi All,

Over the past year we have had a recurring problems with our H&E's.  The come 
out of our Gemini Stainer stripped but not on every run; we can go for months 
without a problem and then all of a sudden it is back again.  The stripping is 
in the Eosin, our pathologist confirms the the hemotoxylin is not affected.  
There are 3 of us cutting on 3 different microtomes.  The age on the microtomes 
range in age from 10 years to less than 1 year. 

Some of the solution we have tried are:

We numbered all our racks to see if some of our older racks may have micro 
cracks in them that may retain solution which gradually dripped onto the 
slides.  After checking for over a month there was no pattern to  racks and bad 
staining.

We tried lowering the level of the solutions to below the coated end of the 
slides.  No results

We tried changing the positions of the pots for the solutions,  thinking that 
if there was a leak from the solutions above we would notice a difference.  No 
luck.

We checked to see if the stripes only appeared near the end of our stains life. 
 Not that either

Right now I have a technician sitting in front of the stainer as it runs to see 
if she can catch any abnormalities in the way the machine is transferring the 
slides.

We rotate our alcohols every run.

We change every solution after every 450 slides.

I have put a call into the company to see if I can get to speak to a specialist 
on this machine but in the mean time I was hoping someone out there could help. 
 Has anyone seen such a problem

I, we, have run out of ideas aside from it being the machine and not being able 
to find out exactly how.


Jo-Ann Bader
Histology Co-Ordinator
Goodman Cancer Centre
McGill University 1160 Pine Ave. W - Rm 312 (3355)
Montreal, QC, Canada
H3G 1Y6
Tel: 514-398-8270
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Re: [Histonet] pax2

[Angela Bitting]

CC1 standard, 32 min incubation with heat disabled. I use Cellmarques predilute
 and UltraView DAB detection.

>>> Dorothy Glass  7/7/2011 3:28 PM >>>
Does anyone have a working protocol for pax2 on the Ventana Ultra or XT?  Can 
you please share on Histonet?

Dorothy Glass
Supervisor, SEPA LABS
Brunswick,Ga.
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IMPORTANT WARNING: The information in this message (and the documents attached 
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prohibited and may be unlawful. If you have received this message in error, 
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[Histonet] pax2

[Dorothy Glass]

Does anyone have a working protocol for pax2 on the Ventana Ultra or XT?  Can 
you please share on Histonet?
 
Dorothy Glass
Supervisor, SEPA LABS
Brunswick,Ga.
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[Histonet] Specimen Collection

[Elaine]

You DO have to tell physicians exactly how to collect specimens. They 
know how to do their job but they have no idea how we do ours, even if 
they did a rotation thru pathology. Sometimes, you may have to tell them 
each time they want to send a specimen if they don't do it routinely. We 
still have staff that call each time they need to send a muscle bx even 
though they've been told numerous times and have received written 
instructions.
Don't think of it as telling them how to do their job, but rather 
consider it good patient care for wanting to make sure it's done right 
the first time.


Elaine
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[Histonet] Specimen Collection Instructions

[Mike Pence]

How has everyone handled this  question GEN.40104
Instructions are distributed to physicians and paramedical personnel for
proper collection, handling, transportation, and preparation of
cytologic and tissue specimens.
 
I feel this question is asking me to spell out to a Dr how to collect
his/her specimens. To me this is like telling him/her how to practice
medicine.
 
I hope someone would like to share what you have seen that works.
 
Thanks, Mike
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[Histonet] Bond III

[Nails, Felton]

We have been using the Bond III for about 4 months and have starting 
experiencing staining on our negative control slide.
We inserted and additional blocking solution and it help for the IHC stains but 
we are still experiencing the problem the with insitu.
Has anyone experienced this problem before and how did you solve it? 

Thanks, Felton
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RE: [Histonet] Request to help for rat eye histology

[CHRISTIE GOWAN]

Are you fixing the eyes in Davidson's? We found that using this fixative which 
I would have to look up since it has been so long since I did rat eyes helped 
maintain better morphology.
 

> From: himanshu.gu...@ucdenver.edu
> To: histonet@lists.utsouthwestern.edu
> Date: Thu, 7 Jul 2011 11:37:39 -0600
> Subject: [Histonet] Request to help for rat eye histology
> 
> Hi
> 
> I need an advice from all experts there on histology of Rat eye.
> 
> I need to take clear section of eye (cross section) so that I can see each 
> layer of cells clearly. Please advise me step by step process to prepare the 
> histology slides/sections of rat eye.
> 
> I try it many times but I am not able to get the clear sections, the 
> cells/layers got broken.
> 
> Regards
> 
> himanshu
> 
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[Histonet] RE: Request to help for rat eye histology - long response

[Elizabeth Chlipala]

We process rat eyes all of the time.  I know that the collection of the eyes is 
important and I can not comment on that since we do not collect the eyes, but 
anytime you are handling both mouse or rat eyes for retinal work you need to be 
as gentle as possible.  For retinal work you need to fix in davidsons fixative 
(don't know how long, since we receive the eyes in 70% alcohol) formalin 
fixative will not work the retina will detach.  One we receive the eyes which 
have been marked with ink as to the superior portion, we trim about 1mm or so 
on the nasal side (you will also need to know if the eye is right or left to 
help you determine which is the nasal side).  I trim with a microtome blade and 
using a small round trimming matrix that the eye sits on top.  The eyes are 
processed on a same day processing cycle - 20 minutes per station with proper 
instead of xylene.  We do not process overnight.

70% - 20 minutes
80% - 20 minutes
95% - 20 minutes
100% - 20 minutes
100% - 20 minutes
Propar - 20 minutes
Propar - 20 minutes
Propar - 20 minutes
Paraffin - 20 minutes
Paraffin - 20 minutes
Paraffin - 20 minutes

When embedding we have a deep base mold filled with melted paraffin that we 
drop the eye into to make sure that there are no air bubbles in the eye 
caviety.  You can gently move the lens to get the air bubbles out if necessary, 
but you really do not want to manipulate the eye much since you might cause 
separation and tearing of the structures. Once we know that the eye has no air 
bubbles we place it in the appropriate base mold, we use the small square molds 
with the superior side of the eye towards the cassette label.  We trim into the 
correct area, we have a microscope next to the microtome so we can make sure 
that we are at the optic nerve head and then place the trimmed blocks on ice 
for about 30 minutes or so, we then cut ribbons of 5 micron sections depending 
upon the study design and place on a good plus slide, let dry overnight and 
stain the following day with H&E.  Since you are in Denver we are just in 
Longmont so if you want to stop by and see how we do this just e-mail me back.

Good Luck

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
www.premierlab.com

Ship to address:

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 Gupta, Himanshu
Sent: Thursday, July 07, 2011 11:38 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Request to help for rat eye histology

Hi

I need an advice from all experts there on histology of Rat eye.

I need to take clear section of eye (cross section) so that I can see each 
layer of cells clearly. Please advise me step by step process to prepare the 
histology slides/sections of rat eye.

I try it many times but I am not able to get the clear sections, the 
cells/layers got broken.

Regards

himanshu

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[Histonet] Request to help for rat eye histology

[Gupta, Himanshu]

Hi

I need an advice from all experts there on histology of Rat eye.

I need to take clear section of eye (cross section) so that I can see each 
layer of cells clearly. Please advise me step by step process to prepare the 
histology slides/sections of rat eye.

I try it many times but I am not able to get the clear sections, the 
cells/layers got broken.

Regards

himanshu

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RE: [Histonet] RE: Tissue Cassette Printer Recommendations

[WILLIAM DESALVO]

I agree with the previous comments and we are in the process of installing 
Vantage. During our testing we found that Ventana will work with any vendor, 
the key issue is interfacing to your LIS.  As part of the process, we tested 
several vendor cassette printers. We chose Thermo's new Printmate 150 for a 
combination of reasons that worked for our process; tape transfer, speed of 
print, footprint (using at the grossing station) and quality of print (2D and 
Linear).
 
Thermo has a 2 hopper and 6 hopper Printmate, heat transfer and prints 
cassettes with 2D bar code in about 8 sec
General Data sells the the 1 hopper CL-01 and 12 hopper CL-12. Laser process 
that requires a special coated cassette that allows the Laser to etch. Prints 
in about 4 sec
Leica sells the CL-01 and their older model IPC, inkjet and special treatment 
of print.
Sakura sells a similar model of the Leica IPC
 
Consider using "light" colored cassettes. The darker and red colored are more 
difficult to get the hand help bar code readers to recognize (red laser).
 
Contact the vendors and I am sure they will be glad to let you demo their 
instrument for several days. Test them all and decide what works best in your 
workflow and process.


William DeSalvo, B.S., HTL(ASCP)

 

> From: jel...@yumaregional.org
> To: cb...@lexclin.com; l...@premierlab.com; arodrig...@emc.org; 
> histonet@lists.utsouthwestern.edu
> Date: Thu, 7 Jul 2011 17:05:11 +
> CC: 
> Subject: [Histonet] RE: Tissue Cassette Printer Recommendations
> 
> I do not have Vantage, so I cannot comment on that. But you have to take into 
> account the environment and the scanning quality of the print. I would also 
> look at space, location, downtime, workflow, even clarity of barcode meaning 
> Datamatirx, ect. There are also issues with specific cassette colors and 
> cassette surface texture playing a part in the scanning.
> 
> -Original Message-
> From: Carol Bryant [mailto:cb...@lexclin.com] 
> Sent: Thursday, July 07, 2011 9:55 AM
> To: 'Elizabeth Chlipala'; Jesus Ellin; 'Rodriguez, Arnold'; 
> histonet@lists.utsouthwestern.edu
> Subject: RE: Tissue Cassette Printer Recommendations
> 
> I am intersted in this info also. I would like to know what printers everyone 
> are using with the Vantage system or a bar coding system. 
> Thank you,
> Carol 
> 
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu 
> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth 
> Chlipala
> Sent: Thursday, July 07, 2011 12:53 PM
> To: 'Jesus Ellin'; 'Rodriguez, Arnold'; histonet@lists.utsouthwestern.edu
> Subject: [Histonet] RE: Tissue Cassette Printer Recommendations
> 
> I second Jesus's comments especially if you are moving towards bar coding. 
> You will need a cassette printer that can print a high quality 2D bar code.
> 
> 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
> www.premierlab.com
> 
> Ship to address:
> 
> 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 Jesus Ellin
> Sent: Thursday, July 07, 2011 10:49 AM
> To: 'Rodriguez, Arnold'; histonet@lists.utsouthwestern.edu
> Subject: [Histonet] RE: Tissue Cassette Printer Recommendations
> 
> I would look at Thermo, Leica and General Data printers .
> 
> Jesus Ellin
> Yuma Regional Medical Center
> 
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu 
> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rodriguez, 
> Arnold
> Sent: Thursday, July 07, 2011 9:38 AM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Tissue Cassette Printer Recommendations
> 
> Hello All,
> 
> We are interested in purchasing a new cassette printer and I would
> sincerely appreciate any recommendations for this instrument. We are
> particularly looking for reliability, print speed, LIS connectivity and
> barcode technology.
> 
> Thank you very much.
> 
> Arnold Rodriguez, HT (ASCP)
> Supervisor, Anatomic Pathology
> Eisenhower Medical Center
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> 
> 
> DISCLAIMER:
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> may contain information that is privileged or exempt from disclosure under 
> applicable law. If you are not the intended recipient(s), you are notified 
> that the dissemination, distribution, or copying of this message is strictly 
> prohibited, if you receive this message in error, or are not the named 
> recipient(s), please notify the sender at either the e-mail, fax, address, or 
> telephone number listed above and delete this e-mail from your compu

RE: [Histonet] RE: Tissue Cassette Printer Recommendations

[CHRISTIE GOWAN]

 The Sakura and the Leica are the same system as they share a patent (this is 
what I am told). We use the Leica IPC for our cassettes and we have the Vantage 
system. We did a demo of the Thermo printer but were not happy with the speed 
and the fact that we would need to change who we order cassettes from. We did 
change the color of our routine cassettes from blue to white as the white 
scanned better at the vantage stations. We still use blue but only for autopsy 
cases. We use the Vantage system at the printer to scan in the patient bar 
code, choose a hopper with the color we want and hit print all. So far it has 
worked quite well.
Christie Gowan
cgo...@uabmc.edu
 

  
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[Histonet] RE: Tissue Cassette Printer Recommendations

[Jesus Ellin]

I do not have Vantage, so I cannot comment on that. But you have to take into 
account the environment and the scanning quality of the print.  I would also 
look at space, location, downtime, workflow, even clarity of barcode meaning 
Datamatirx, ect.  There are also issues with specific cassette colors and 
cassette surface texture playing a part in the scanning.

-Original Message-
From: Carol Bryant [mailto:cb...@lexclin.com] 
Sent: Thursday, July 07, 2011 9:55 AM
To: 'Elizabeth Chlipala'; Jesus Ellin; 'Rodriguez, Arnold'; 
histonet@lists.utsouthwestern.edu
Subject: RE: Tissue Cassette Printer Recommendations

I am intersted in this info also.  I would like to know what printers everyone 
are using with the Vantage system or a bar coding system. 
Thank you,
Carol 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth 
Chlipala
Sent: Thursday, July 07, 2011 12:53 PM
To: 'Jesus Ellin'; 'Rodriguez, Arnold'; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Tissue Cassette Printer Recommendations

I second Jesus's comments especially if you are moving towards bar coding.  You 
will need a cassette printer that can print a high quality 2D bar code.

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
www.premierlab.com

Ship to address:

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 Jesus Ellin
Sent: Thursday, July 07, 2011 10:49 AM
To: 'Rodriguez, Arnold'; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Tissue Cassette Printer Recommendations

I would look at Thermo, Leica and General Data printers .

Jesus Ellin
Yuma Regional Medical Center

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rodriguez, 
Arnold
Sent: Thursday, July 07, 2011 9:38 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Tissue Cassette Printer Recommendations

Hello All,

We are interested in purchasing a new cassette printer and I would
sincerely appreciate any recommendations for this instrument. We are
particularly looking for reliability, print speed, LIS connectivity and
barcode technology.

Thank you very much.

Arnold Rodriguez, HT (ASCP)
Supervisor, Anatomic Pathology
Eisenhower Medical Center
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you.

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[Histonet] RE: Tissue Cassette Printer Recommendations

[Weems, Joyce]

We have used Sakura for years now --- cassette and slide. They do a great job.  


Joyce Weems 
Pathology Manager 
Saint Joseph's Hospital 
5665 Peachtree Dunwoody Rd NE 
Atlanta, GA 30342 
678-843-7376 - Phone 
678-843-7831 - Fax 


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rodriguez, 
Arnold
Sent: Thursday, July 07, 2011 12:38
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Tissue Cassette Printer Recommendations

Hello All,
 
We are interested in purchasing a new cassette printer and I would sincerely 
appreciate any recommendations for this instrument. We are particularly looking 
for reliability, print speed, LIS connectivity and barcode technology.
 
Thank you very much.
 
Arnold Rodriguez, HT (ASCP)
Supervisor, Anatomic Pathology
Eisenhower Medical Center
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[Histonet] RE: Tissue Cassette Printer Recommendations

[Carol Bryant]

I am intersted in this info also.  I would like to know what printers everyone 
are using with the Vantage system or a bar coding system. 
Thank you,
Carol 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth 
Chlipala
Sent: Thursday, July 07, 2011 12:53 PM
To: 'Jesus Ellin'; 'Rodriguez, Arnold'; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Tissue Cassette Printer Recommendations

I second Jesus's comments especially if you are moving towards bar coding.  You 
will need a cassette printer that can print a high quality 2D bar code.

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
www.premierlab.com

Ship to address:

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 Jesus Ellin
Sent: Thursday, July 07, 2011 10:49 AM
To: 'Rodriguez, Arnold'; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Tissue Cassette Printer Recommendations

I would look at Thermo, Leica and General Data printers .

Jesus Ellin
Yuma Regional Medical Center

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rodriguez, 
Arnold
Sent: Thursday, July 07, 2011 9:38 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Tissue Cassette Printer Recommendations

Hello All,

We are interested in purchasing a new cassette printer and I would
sincerely appreciate any recommendations for this instrument. We are
particularly looking for reliability, print speed, LIS connectivity and
barcode technology.

Thank you very much.

Arnold Rodriguez, HT (ASCP)
Supervisor, Anatomic Pathology
Eisenhower Medical Center
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[Histonet] RE: Tissue Cassette Printer Recommendations

[Elizabeth Chlipala]

I second Jesus's comments especially if you are moving towards bar coding.  You 
will need a cassette printer that can print a high quality 2D bar code.

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
www.premierlab.com

Ship to address:

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 Jesus Ellin
Sent: Thursday, July 07, 2011 10:49 AM
To: 'Rodriguez, Arnold'; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Tissue Cassette Printer Recommendations

I would look at Thermo, Leica and General Data printers .

Jesus Ellin
Yuma Regional Medical Center

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rodriguez, 
Arnold
Sent: Thursday, July 07, 2011 9:38 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Tissue Cassette Printer Recommendations

Hello All,

We are interested in purchasing a new cassette printer and I would
sincerely appreciate any recommendations for this instrument. We are
particularly looking for reliability, print speed, LIS connectivity and
barcode technology.

Thank you very much.

Arnold Rodriguez, HT (ASCP)
Supervisor, Anatomic Pathology
Eisenhower Medical Center
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you.

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[Histonet] RE: Tissue Cassette Printer Recommendations

[Jesus Ellin]

I would look at Thermo, Leica and General Data printers .

Jesus Ellin
Yuma Regional Medical Center

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rodriguez, 
Arnold
Sent: Thursday, July 07, 2011 9:38 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Tissue Cassette Printer Recommendations

Hello All,
 
We are interested in purchasing a new cassette printer and I would
sincerely appreciate any recommendations for this instrument. We are
particularly looking for reliability, print speed, LIS connectivity and
barcode technology.
 
Thank you very much.
 
Arnold Rodriguez, HT (ASCP)
Supervisor, Anatomic Pathology
Eisenhower Medical Center 
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prohibited, if you receive this message in error, or are not the named 
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telephone number listed above and delete this e-mail from your computer.  Thank 
you.

__
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Thank You.
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[Histonet] Bone Marrows

[Marcia Fisher]

We assist with bone marrows either at bedside or in Outpatient Observation, 
never in the OR.  All other bone marrows are performed at the Oncologist's 
office.

Marcia Fisher
Histology Supervisor/Lab Safety Officer
El Centro Regional Medical Center
1415 Ross Ave
El Centro, CA  92243
760-339-7267
760-482-5365(F)
www.ecrmc.org

Confidentiality Notice: This e-mail is for the sole use of the intended 
recipient(s) and may contain confidential and privileged information. Any 
unauthorized review, use, disclosure, or distribution is prohibited. If you are 
not the intended recipient, please contact the sender at the phone number above 
and promptly destroy this e-mail and its attachments.



ECRMC Confidentiality Notice: This e-mail is for the sole use of the intended 
recipient(s) and may contain confidential and privileged information. Any 
unauthorized review, use, disclosure, or distribution is prohibited. If you are 
not the intended recipient, PLEASE contact the sender and promptly destroy this 
e-mail and its attachments.
 


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[Histonet] Tissue Cassette Printer Recommendations

[Rodriguez, Arnold]

Hello All,
 
We are interested in purchasing a new cassette printer and I would
sincerely appreciate any recommendations for this instrument. We are
particularly looking for reliability, print speed, LIS connectivity and
barcode technology.
 
Thank you very much.
 
Arnold Rodriguez, HT (ASCP)
Supervisor, Anatomic Pathology
Eisenhower Medical Center 
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RE: [Histonet] Routine H & E stain,

[Rittman, Barry R]

I believe that this could be a multifactorial problem.
There is a tendency for us, "in the interests of efficiency" to process tissue 
and slides as rapidly as possible. In a lot of cases I suspect that the 
absolute minimum time in reagents causes some carry over that can result in 
fading of the stains.
The quality of the xylene, as one person already pointed out may be a factor. 
Even using "real xylene", the composition, sulfur content etc can vary 
considerably and affect staining. There was some concern in the 1950s over 
fading of slides containing silver stains due to using xylene. However I still 
have some stained with silver that were cleared with xylene and still look 
really good.
The use of xylene substitutes has not (to my admittedly imperfect knowledge) 
been tested over a few decades.

It would also be interesting to know whether the type of hematoxylin stain used 
has any effect on fading of hematoxylin. I have always been an advocate for 
Ehrlich's hematoxylin and have 50+ year old slides that  are still as bright as 
the day they were stained but not much experience with other hematoxylins.

Mounting media may also have an effect. In the days of the dinosaur we used 
Canada balsam in xylene and later, DPX. Slides mounted in these still look 
really good, in fact dare I say beautiful.

Might be an idea to use some pH paper to check pH of the xylene and mountants.

Some have also suggested that exposure to UV light may also be a factor. Not 
sure that there is much evidence for this.

I have seen some slides from the mid 1800s that while not pristine still have 
some hematoxylin staining. 

Bottom line, is that we have no bloody idea what is causing your hematoxylin to 
fade- suggest that failing all else, you pray and enjoy a glass of Zinfandel 
(Paso Robles 2006 was a really good year).
Barry

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Thursday, July 07, 2011 9:45 AM
To: Histonet; histonet-boun...@lists.utsouthwestern.edu; histonet; 
amitapan...@torrentpharma.com
Subject: Re: [Histonet] Routine H & E stain,

It seems that your mounting medium is acid. Try to correct that rather than 
changing the staining.
René J.

--- On Thu, 7/7/11, amitapan...@torrentpharma.com 
 wrote:


From: amitapan...@torrentpharma.com 
Subject: [Histonet] Routine H & E stain,
To: "Histonet" , 
histonet-boun...@lists.utsouthwestern.edu, "histonet" 

Date: Thursday, July 7, 2011, 12:22 AM


Hello Histonetters,

I am from toxicopathology lab where we perform H&E on rat tissues and 
store these slides for 10 long years.

We use self lab prepare Harris hematoxylin (Hematoxylin crystal+Alcohol+ 
Ammonium alum +D/W and Mercuric oxide for ripening) and 1% aqueous eosin. 
The staining result is good (Purplish pink look stained slide) on all 
tissue, but  our observation is after 5-6 months time this get faded and 
become towards pinkish type though we can observe the slide. 
We want to retain its fresh stained color. 

Please suggest me how to keep stable stain for longer period or do you 
suggest to switch to any other type of haematoxylin - prepared or 
commercially available?

Looking forward for your feed back.
Amita
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[Histonet] EM supervisor postion at UC San Francisco, job description

[Morken, Timothy]

Here is the EM job  I recently mentioned would be open. See the HR website at   
http://jobs.ucsfmedicalcenter.org/

Job Description



Job ID:











2257



Job Title:



Electron Microscopy Lab Supervisor (HISTOTECHNOLOGIST, SUPVR)



Job Code:



9068



Department:



Pathology-Surgical / Histology



Location:

Parnassus



Full/Part Time:















Full-Time



Regular/Temporary:



Regular



Shift:



Not Applicable



Weekly Hours:



40



Salary Range:



$68993.6 - $113776



Union Information:





This classification is not represented by a union





























Return to Previous 
Page

















Appointment Type: Career

Percentage:  100%

Shift: Days

Shift Length: 8 hours

Work Days:  Monday-Friday

Department: Pathology

Summary of Duties:
Under general direction from the Director of Clinical Operations and Medical 
Director, the incumbent serves as supervisor to 3 Histotechnologists.   In 
addition to supervisory duties, the incumbent serves as a technical expert 
providing direction to staff as well as performing all technical aspects of 
diagnostic electron microscopy, immunofluorescence microscopy and muscle 
histochemistry procedures.  As supervisor, incumbent recruits, hires, trains, 
completes performance evaluations, and resolves employee issues.  Provides 
orientation, completes competency assessments, maintains staff schedules, 
training and compliance documentation.  Implements new procedures as needed, 
updates and maintains the lab manual and annual reviews required for 
accreditation.  Ensures that all equipment is well-maintained and that staff 
training and documentation are completed as required.  Ensures that QC 
documentation is complete and quality standards are maintained in the 
laboratory.  Advises and assists researchers planning research projects and 
determines the ability of the laboratory to accommodate research projects, 
updating the Medical Director as required.  Other duties as assigned.

Required Qualifications:
* College degree in a biological science, chemistry or a related field 
or equivalent education and work experience in electron microscopy required, 
plus three years of senior-level experience in a hospital pathology laboratory 
performing all aspects of diagnostic electron microscopy work-up.
* Extensive knowledge of tissue ultrastructure, including 
Ultrastructural pathology required.
* Knowledge / demonstrated skill in of electron microscope maintenance 
required.
* Excellent interpersonal communication skills required.
* Demonstrated ability to organize and prioritize responsibilities and 
perform well under pressure to meet deadlines required.
* Previous, recent supervisory experience in an Electron Microscopy 
Laboratory required.
* Understanding of regulatory requirements required.

Preferred Qualifications:
* EMSA Certification in Electron Microscopy strongly preferred.
* Experience in muscle histochemistry highly desirable.
* Formal training in electron microscopy.
* Experience with digital EM imaging.
* Experience in Immunofluorescence technique.
* Laboratory supervisory experience in teaching hospital.

Required Licenses/Certifications:
* N/A







Tim Morken
Supervisor, Histology, IPOX
UC San Francisco Medical Center
Box 1656
1600 Divisidero St, B217
San Francisco, CA 94115
USA

415.514.6042 (office)
415.885.7409 Fax
tim.mor...@ucsfmedctr.org


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Re: [Histonet] Leaving sample over night in ETOH 70% during fixation

[Rene J Buesa]

Bouin's and PFA are completely different in their action an results.
If you get good results with Bouin's use it and try to overcome your cutting 
difficulties.
There should be no differences using PFA or 37% formaldehyde. Methanol is only 
used to prevent formaldehyde polymerization and does not interfere with 
fixation. The problem is that you will need longer fixation times to obtain 
good results when using formaldehyde.René J.

--- On Thu, 7/7/11, Itai Moshe  wrote:



From: Itai Moshe 
Subject: Re: [Histonet] Leaving sample over night in ETOH 70% during fixation
To: "Edwards, Richard E." , 
histonet@lists.utsouthwestern.edu
Date: Thursday, July 7, 2011, 5:09 AM


I had a problem with liver fixation with PFA, most of the time I've got no
signal, so I've switched to Bouin's, and got good resalts, although it is
harder to cut bouin's sections with microtom.

Could it be because that I've switched from Paraformaldehyde powder to
formaldehyde solution (37%) (that contains methanol as a preservative) ?
I have one good PFA  powder experiment that I've done, and
some excellent old experiments that someone else in my lab did many years
ago, but i don't know how, all worked great with PFA fixation, but i could
not replicate the that.

I've tried to do a Formaldehyde 24Hr fixation R.T for the mouse liver
samples, but the results was catastrophic and even the DAPI didn't worked.

So i thought that maybe that because that the working samples were in 70%
ETOH over night.


2011/7/7 Edwards, Richard E. 

> Leaving  tissue overnight in 70% ethanol at 4C should have no detrimental
> effect n the tissues, why 4C overnight with Bouin's??
>
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu [mailto:
> histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Itai Moshe
> Sent: 07 July 2011 09:22
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Leaving sample over night in ETOH 70% during fixation
>
> Dear All,
> Does leaving the samples (diaphragm, liver) over night in ethanol 70% at 4C
> during the fixation process, will be better for the tissue fixation, and
> does not harm the sample ?
> Does the fixation process should be done straight forward from the first
> step to the last one without any over night stops (except from the PFA,
> Bouin's step) ?
>
>
> My fixation protocol is like this:
> 1) immediately after killing the mouse i'm putting the sections in a
> fixation solution that is made from: 10ml formaldehyde 37%+5ml PBSx20+85ml
> DDW  - pH 7, Or bouin's solution over night at 4C.
> 2) ETOH 70%, ETOH 80%, ETOH 96%, ETOH 100% x2 - each for 1Hr at RT
> 3) Xylen x2 - each for 1Hr at RT.
> 4) Paraffin x3 - each at 60C for 1Hr.
>
> Thank you all very much in advance
>
> Itai M
> ___
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> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>


-- 
Itai Moshe
Mark Pines lab
Institute of Animal Sciences, Volcani Center.
Dept. of Animal Sciences, School of Veterinary Medicine, The Hebrew
University of Jerusalem
Israel
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Re: [Histonet] Leaving sample over night in ETOH 70% during fixation

[Rene J Buesa]

If you have an overnight step it is better to prolong the formalin fixation 
instead of leaving the tissues in 70EthOL.
Since formalin will require at least 24 hours to completely bind and 48 to 
crosslink, if you leave your tissues overnight in 70EthOL the unfixed tissue 
will be fixed by the alcohol instead and that can cause difficulties in the 
subsequent tests.
René J.

--- On Thu, 7/7/11, Itai Moshe  wrote:


From: Itai Moshe 
Subject: [Histonet] Leaving sample over night in ETOH 70% during fixation
To: histonet@lists.utsouthwestern.edu
Date: Thursday, July 7, 2011, 4:22 AM


Dear All,
Does leaving the samples (diaphragm, liver) over night in ethanol 70% at 4C
during the fixation process, will be better for the tissue fixation, and
does not harm the sample ?
Does the fixation process should be done straight forward from the first
step to the last one without any over night stops (except from the PFA,
Bouin's step) ?


My fixation protocol is like this:
1) immediately after killing the mouse i'm putting the sections in a
fixation solution that is made from: 10ml formaldehyde 37%+5ml PBSx20+85ml
DDW  - pH 7, Or bouin's solution over night at 4C.
2) ETOH 70%, ETOH 80%, ETOH 96%, ETOH 100% x2 - each for 1Hr at RT
3) Xylen x2 - each for 1Hr at RT.
4) Paraffin x3 - each at 60C for 1Hr.

Thank you all very much in advance

Itai M
Doe
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Re: [Histonet] Routine H & E stain,

[Rene J Buesa]

It seems that your mounting medium is acid. Try to correct that rather than 
changing the staining.
René J.

--- On Thu, 7/7/11, amitapan...@torrentpharma.com 
 wrote:


From: amitapan...@torrentpharma.com 
Subject: [Histonet] Routine H & E stain,
To: "Histonet" , 
histonet-boun...@lists.utsouthwestern.edu, "histonet" 

Date: Thursday, July 7, 2011, 12:22 AM


Hello Histonetters,

I am from toxicopathology lab where we perform H&E on rat tissues and 
store these slides for 10 long years.

We use self lab prepare Harris hematoxylin (Hematoxylin crystal+Alcohol+ 
Ammonium alum +D/W and Mercuric oxide for ripening) and 1% aqueous eosin. 
The staining result is good (Purplish pink look stained slide) on all 
tissue, but  our observation is after 5-6 months time this get faded and 
become towards pinkish type though we can observe the slide. 
We want to retain its fresh stained color. 

Please suggest me how to keep stable stain for longer period or do you 
suggest to switch to any other type of haematoxylin - prepared or 
commercially available?

Looking forward for your feed back.
Amita
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Re: [Histonet] Routine H & E stain,

[kgrobert]

Amita,

We are a neurotoxicology lab that also provides histology services for
other researchers inside & outside Rutgers, and we use Gill's #3
hematoxylin for our routine H&Es on mouse and rat tissues.  No fading that
I have noticed in all the years that I have been here.  We get ours from
Thermo Fisher, or whatever they call themselves nowadays.  :o)

Good luck,
Kathleen Roberts

Principal Lab Technician
Neurotoxicology Labs
Molecular Pathology Facility Core
Dept of Pharmacology & Toxicology
Rutgers, the State University of NJ
41 B Gordon Road
Piscataway, NJ 08854
(732) 445-6914


> Hello Histonetters,
>
> I am from toxicopathology lab where we perform H&E on rat tissues and
> store these slides for 10 long years.
>
> We use self lab prepare Harris hematoxylin (Hematoxylin crystal+Alcohol+
> Ammonium alum +D/W and Mercuric oxide for ripening) and 1% aqueous eosin.
> The staining result is good (Purplish pink look stained slide) on all
> tissue, but  our observation is after 5-6 months time this get faded and
> become towards pinkish type though we can observe the slide.
> We want to retain its fresh stained color.
>
> Please suggest me how to keep stable stain for longer period or do you
> suggest to switch to any other type of haematoxylin - prepared or
> commercially available?
>
> Looking forward for your feed back.
> Amita
> ___
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> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>




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Re: [Histonet] Bone Marrows

[DKBoyd]

Yes we do.  We assist with all bone marrows regardles of where they are 
(not hematology).   We dress out in scrubs, etc.

Debbie M. Boyd, HT(ASCP) l Chief Histologist l Southside Regional Medical 
Center I 
200 Medical Park Boulevard l Petersburg, Va.  23805 l T: 804-765-5050 l F: 
804-765-5582 l dkb...@chs.net





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Re: [Histonet] Routine H & E stain,

[SHANE NELSON]

I should also have added not all synthetic Xylenes do this and I also use a 
Microwave Processor. These variables may have been a factor. Wonderful world of 
Histology, TRIAL AND ERROR. 
 
THANK YOU,
 
PATTI RUBEN-NELSON  H.T.(ASCP) 
PNP LABORATORY CONSULTANTS
SUPERVISOR/DGC
P.O. BOX 412
CABAZON, CA. 92230
cell (909) 841-9761 
nelsonr...@verizon.net





From: "amitapan...@torrentpharma.com" 
To: Histonet ; 
histonet-boun...@lists.utsouthwestern.edu; histonet 

Sent: Wed, July 6, 2011 9:22:38 PM
Subject: [Histonet] Routine H & E stain,

Hello Histonetters,

I am from toxicopathology lab where we perform H&E on rat tissues and 
store these slides for 10 long years.

We use self lab prepare Harris hematoxylin (Hematoxylin crystal+Alcohol+ 
Ammonium alum +D/W and Mercuric oxide for ripening) and 1% aqueous eosin. 
The staining result is good (Purplish pink look stained slide) on all 
tissue, but  our observation is after 5-6 months time this get faded and 
become towards pinkish type though we can observe the slide. 
We want to retain its fresh stained color. 

Please suggest me how to keep stable stain for longer period or do you 
suggest to switch to any other type of haematoxylin - prepared or 
commercially available?

Looking forward for your feed back.
Amita
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Re: [Histonet] Routine H & E stain,

[SHANE NELSON]

Are you using synthetic xylene in your H&E/COVER SLIPPING protocol. If yes, you 
might want to consider ending your protocol with the real thing (XYLENE). I had 
the same situation until we switched to real XYLENE. No more H&E fading.




THANK YOU,
 
PATTI RUBEN-NELSON  H.T.(ASCP) 
PNP LABORATORY CONSULTANTS
SUPERVISOR/DGC
P.O. BOX 412
CABAZON, CA. 92230
cell (909) 841-9761 
nelsonr...@verizon.net





From: "amitapan...@torrentpharma.com" 
To: Histonet ; 
histonet-boun...@lists.utsouthwestern.edu; histonet 

Sent: Wed, July 6, 2011 9:22:38 PM
Subject: [Histonet] Routine H & E stain,

Hello Histonetters,

I am from toxicopathology lab where we perform H&E on rat tissues and 
store these slides for 10 long years.

We use self lab prepare Harris hematoxylin (Hematoxylin crystal+Alcohol+ 
Ammonium alum +D/W and Mercuric oxide for ripening) and 1% aqueous eosin. 
The staining result is good (Purplish pink look stained slide) on all 
tissue, but  our observation is after 5-6 months time this get faded and 
become towards pinkish type though we can observe the slide. 
We want to retain its fresh stained color. 

Please suggest me how to keep stable stain for longer period or do you 
suggest to switch to any other type of haematoxylin - prepared or 
commercially available?

Looking forward for your feed back.
Amita
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[Histonet] RE: Bone Marrows

[Clare Thornton]

I do, as well as several of my coworkers.  We assist with bone marrows in 
inpatient rooms, pediatric sedation, outpatient surgery center, and 
occasionally in the main OR.  What is your question?


Clare J. Thornton, HTL(ASCP)
Assistant Histology Supervisor
Dahl-Chase Diagnostic Services
417 State Street, Suite 540
Bangor, ME 04401
cthorn...@dahlchase.com

From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kennedy, Lisa 
[lisakenn...@catholichealth.net]
Sent: Thursday, July 07, 2011 5:52 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Bone Marrows

Are there any Histo Techs out there who assist with bone marrows IN the
operating room?
Lisa
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[Histonet] Bone Marrows

[Kennedy, Lisa]

Are there any Histo Techs out there who assist with bone marrows IN the
operating room?
Lisa
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RE: [Histonet] Leaving sample over night in ETOH 70% during fixation

[Margaret Blount]

Storing your samples overnight in 70% ethanol after fixation won't harm your 
samples, but they should be adequately fixed prior to this. I do not like 
xylene for mouse tissues, it makes them too brittle. Instead I used histoclear 
11 from National Diagnostics. Histoclear 11 is less expensive than the original 
histoclear, but performs well in my experience. I found that for most of the 
tissues I processed (I never tried diaphragm) a shorter schedule worked well 
and in fact the process you describe may be too long for some tissues. The 
Society of histotechnologists produces a booklet with suggested processing 
schedules for a wide range of animals and tissues, I found this invaluable in 
designing my protocols. My process for mouse tissues took aroung 6 hours using 
a processing machine equipped with vacuum on all stations. Unfortunately as I 
have retired I don't have access to my SOP's any more. However, if you obtain 
the booklet from the Society, you will be able to devise a good protocol. At 
the end of the day, if your tissues section and stain well, then your process 
is satisfactory. 
It's always helpful to compare your sections with other people's if you can 
then you have a benchmark of quality. This can be difficult and frustrating in 
a small research lab. 
 
You may find that postfixing your liver samples in formol alcohol (90ml 
absolute ethanol to 10ml 37% formaldehyde or use Pen fix from thermo shandon) 
will improve the morphology. I used to do this for a couple of hours; you may 
need to experiment, especially if you are doing immunohistochemistry which is 
presumably why you are using paraformaldehyde. 

 

Good luck

 

Margaret




From: histonet-boun...@lists.utsouthwestern.edu on behalf of Itai Moshe
Sent: Thu 07/07/2011 09:22
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Leaving sample over night in ETOH 70% during fixation



Dear All,
Does leaving the samples (diaphragm, liver) over night in ethanol 70% at 4C
during the fixation process, will be better for the tissue fixation, and
does not harm the sample ?
Does the fixation process should be done straight forward from the first
step to the last one without any over night stops (except from the PFA,
Bouin's step) ?


My fixation protocol is like this:
1) immediately after killing the mouse i'm putting the sections in a
fixation solution that is made from: 10ml formaldehyde 37%+5ml PBSx20+85ml
DDW  - pH 7, Or bouin's solution over night at 4C.
2) ETOH 70%, ETOH 80%, ETOH 96%, ETOH 100% x2 - each for 1Hr at RT
3) Xylen x2 - each for 1Hr at RT.
4) Paraffin x3 - each at 60C for 1Hr.

Thank you all very much in advance

Itai M
Doe
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Re: [Histonet] Leaving sample over night in ETOH 70% during fixation

[Itai Moshe]

Thank's,

Did you used it for mouse liver section ?
Can you please post a your fixation protocol ?

2011/7/7 Edwards, Richard E. 

>  I  have no  experience of using  Bouin’s fixed tissue  for  immunos, tho’
> it  is an excellent fixative for many tinctorial procedures providing the
> fixation time is carefully  controlled…I do not think that  the  methanol in
> the  formalin could affect  anything adversely, also  10% neutral buffered
> formalin is the  fixative  of choice  for  most  labs, I  have never  had
> a  problem with it.
>
> ** **
>
> *From:* Itai Moshe [mailto:itai.mo...@mail.huji.ac.il]
> *Sent:* 07 July 2011 10:10
> *To:* Edwards, Richard E.; histonet@lists.utsouthwestern.edu
> *Subject:* Re: [Histonet] Leaving sample over night in ETOH 70% during
> fixation
>
> ** **
>
> I had a problem with liver fixation with PFA, most of the time I've got no
> signal, so I've switched to Bouin's, and got good resalts, although it is
> harder to cut bouin's sections with microtom.
>
> ** **
>
> Could it be because that I've switched from Paraformaldehyde powder to
> formaldehyde solution (37%) (that contains methanol as a preservative) ?**
> **
>
> I have one good PFA  powder experiment that I've done, and
> some excellent old experiments that someone else in my lab did many years
> ago, but i don't know how, all worked great with PFA fixation, but i could
> not replicate the that.
>
> ** **
>
> I've tried to do a Formaldehyde 24Hr fixation R.T for the mouse liver
> samples, but the results was catastrophic and even the DAPI didn't worked.
> 
>
> ** **
>
> So i thought that maybe that because that the working samples were in 70%
> ETOH over night.
>
> ** **
>
> ** **
>
> 2011/7/7 Edwards, Richard E. 
>
> Leaving  tissue overnight in 70% ethanol at 4C should have no detrimental
> effect n the tissues, why 4C overnight with Bouin's??
>
>
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu [mailto:
> histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Itai Moshe
> Sent: 07 July 2011 09:22
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Leaving sample over night in ETOH 70% during fixation
>
> Dear All,
> Does leaving the samples (diaphragm, liver) over night in ethanol 70% at 4C
> during the fixation process, will be better for the tissue fixation, and
> does not harm the sample ?
> Does the fixation process should be done straight forward from the first
> step to the last one without any over night stops (except from the PFA,
> Bouin's step) ?
>
>
> My fixation protocol is like this:
> 1) immediately after killing the mouse i'm putting the sections in a
> fixation solution that is made from: 10ml formaldehyde 37%+5ml PBSx20+85ml
> DDW  - pH 7, Or bouin's solution over night at 4C.
> 2) ETOH 70%, ETOH 80%, ETOH 96%, ETOH 100% x2 - each for 1Hr at RT
> 3) Xylen x2 - each for 1Hr at RT.
> 4) Paraffin x3 - each at 60C for 1Hr.
>
> Thank you all very much in advance
>
> Itai M
>
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> --
> Itai Moshe
> Mark Pines lab
> Institute of Animal Sciences, Volcani Center.
> Dept. of Animal Sciences, School of Veterinary Medicine, The Hebrew
> University of Jerusalem
> Israel
>


-- 
Itai Moshe
Mark Pines lab
Institute of Animal Sciences, Volcani Center.
Dept. of Animal Sciences, School of Veterinary Medicine, The Hebrew
University of Jerusalem
Israel
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Re: [Histonet] Leaving sample over night in ETOH 70% during fixation

[Itai Moshe]

I had a problem with liver fixation with PFA, most of the time I've got no
signal, so I've switched to Bouin's, and got good resalts, although it is
harder to cut bouin's sections with microtom.

Could it be because that I've switched from Paraformaldehyde powder to
formaldehyde solution (37%) (that contains methanol as a preservative) ?
I have one good PFA  powder experiment that I've done, and
some excellent old experiments that someone else in my lab did many years
ago, but i don't know how, all worked great with PFA fixation, but i could
not replicate the that.

I've tried to do a Formaldehyde 24Hr fixation R.T for the mouse liver
samples, but the results was catastrophic and even the DAPI didn't worked.

So i thought that maybe that because that the working samples were in 70%
ETOH over night.


2011/7/7 Edwards, Richard E. 

> Leaving  tissue overnight in 70% ethanol at 4C should have no detrimental
> effect n the tissues, why 4C overnight with Bouin's??
>
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu [mailto:
> histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Itai Moshe
> Sent: 07 July 2011 09:22
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Leaving sample over night in ETOH 70% during fixation
>
> Dear All,
> Does leaving the samples (diaphragm, liver) over night in ethanol 70% at 4C
> during the fixation process, will be better for the tissue fixation, and
> does not harm the sample ?
> Does the fixation process should be done straight forward from the first
> step to the last one without any over night stops (except from the PFA,
> Bouin's step) ?
>
>
> My fixation protocol is like this:
> 1) immediately after killing the mouse i'm putting the sections in a
> fixation solution that is made from: 10ml formaldehyde 37%+5ml PBSx20+85ml
> DDW  - pH 7, Or bouin's solution over night at 4C.
> 2) ETOH 70%, ETOH 80%, ETOH 96%, ETOH 100% x2 - each for 1Hr at RT
> 3) Xylen x2 - each for 1Hr at RT.
> 4) Paraffin x3 - each at 60C for 1Hr.
>
> Thank you all very much in advance
>
> Itai M
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>


-- 
Itai Moshe
Mark Pines lab
Institute of Animal Sciences, Volcani Center.
Dept. of Animal Sciences, School of Veterinary Medicine, The Hebrew
University of Jerusalem
Israel
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[Histonet] Leaving sample over night in ETOH 70% during fixation

[Itai Moshe]

Dear All,
Does leaving the samples (diaphragm, liver) over night in ethanol 70% at 4C
during the fixation process, will be better for the tissue fixation, and
does not harm the sample ?
Does the fixation process should be done straight forward from the first
step to the last one without any over night stops (except from the PFA,
Bouin's step) ?


My fixation protocol is like this:
1) immediately after killing the mouse i'm putting the sections in a
fixation solution that is made from: 10ml formaldehyde 37%+5ml PBSx20+85ml
DDW  - pH 7, Or bouin's solution over night at 4C.
2) ETOH 70%, ETOH 80%, ETOH 96%, ETOH 100% x2 - each for 1Hr at RT
3) Xylen x2 - each for 1Hr at RT.
4) Paraffin x3 - each at 60C for 1Hr.

Thank you all very much in advance

Itai M
Doe
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