[Histonet] Re: FNA stain

[Robert Richmond]

Debbie M. Boyd, HT(ASCP), Chief Histologist, Southside Regional
Medical Center in Petersburg, Virginia asks:

>>This question is for those of you who perform fine needle aspirations. What 
>>stain are you using for your immediate evaluation?  Or do you give an 
>>immediate evaluation/adequacy?<<

This is a really good question that demands a careful answer. You need
to do what your cytotechnologist and your pathologist are comfortable
with. Air-dried smears stained with a rapid azure-eosin (Diff-Quik and
generic equivalents) stain are simple and fast - IF your pathologist
has been trained to interpret them, which I (I'm 70 years old, mind
you) frankly wasn't. I can live with a good H and E stain - a
full-dress Pap stain is nice, but probably takes too long to do.

The cytotechnologist and pathologist certainly have to give an
immediate decision as to the adequacy of the specimen. There's a CPT
code for that, even. And sometimes what's needed is a flat-out
diagnosis.

But this is something that needs to be worked out in advance - not
under the stress of a high-pressure clinical situation, but
comfortably, with pizza or worse.

Bob Richmond
Samurai Pathologist
Knoxville TN

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Re: [Histonet] looking for KOH nail procedure

[Joe Nocito]

we use 20% sodium hydroxide. Works great
- Original Message - 
From: "Cheryl" 

To: 
Sent: Wednesday, December 16, 2009 4:08 PM
Subject: [Histonet] looking for KOH nail procedure


Hi Guys!

I've used a potassium hydroxide solution to 'soften' nails after fixation 
but before processing. I cannot find my procedure!!


Can anyone help?

Cheryl Kerry, HT(ASCP)

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Re: [Histonet] restaining IHC slides

[Rena Fail]

you can use the same slides , repeating all the steps except  the antigen 
retrieval ( if it is the same ) Ideally you should use new slides, but on tiny 
bxs, outside slides, or very limited area of interest, that is not aways 
possible.
Rena Fail 



- Original Message 
From: Thomas Pier 
To: histonet@lists.utsouthwestern.edu
Sent: Wed, December 16, 2009 1:44:25 PM
Subject: [Histonet] restaining IHC slides

Hello Histonetters,
I ran into an issue when staining some TMA slides the other day. This question 
isn't about how to fix the staining, or lack there of. I would like to know 
whether or not it is possible to repeat the same protocol on the same slides 
after removing the coverslips. If any of you have any experience with this sort 
of thing, please let me know.

Tom Pier
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[Histonet] RE: Bubbles on Fresh Frozen tissue after IHC

[Nick Dee]

 

Hi-

 

I am doing immunohistochemistry on fresh frozen tissue using the
capillary gap method on Tecans.  The preferred fixative for my antigen
is a cold acetone fix.  I am using MeOH/H2O2 to kill peroxidase before
the blocking step and it's resulting in many small bubbles forming on
the tissue in which I get no staining...in the area where there aren't
bubbles, the staining looks great.  I am aware of using something other
than MeOH to block for peroxidase, however when I don't use an alcohol,
the surface tension pulls all the tissue off my slide when I'm taking
apart the capillary gap chambers to coverslip.

 

Has anyone experienced this before?  Any ideas/suggestions would be
appreciated.

 

Thank you, 

Nick 

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RE: [Histonet] immunoperoxidase background staining

[Gomez, Milton]

These are brand new installs.  it does it only for the CC1 and CC2 Protocols.
 
Milton A. Gomez, HTL (ASCP)
Technical Supervisor
Immunohistochemistry Department
ARUP Laboratories, Inc.
500 Chipeta Way
Salt Lake City, UT 84108-1221
Desk Phone:  801-583-2787, ext.3869
Lab. Phone:   801-584-5257/5242
Fax:  801-584-5217
E-mail:  milton.go...@aruplab.com  
Web:  www.aruplab.com  



From: Jackie Smith [mailto:roosmi...@hotmail.com]
Sent: Wed 12/16/2009 3:24 PM
To: Gomez, Milton
Subject: RE: [Histonet] immunoperoxidase background staining



When was the last decontamination performed?  Is it all of your protocols?


 
> Date: Wed, 16 Dec 2009 13:48:51 -0700
> From: milton.go...@aruplab.com
> To: Histonet@lists.utsouthwestern.edu
> CC: 
> Subject: [Histonet] immunoperoxidase background staining
> 
> Dear Histonetters,
> 
> I am getting background staining with my Immunohistochemistry protocols that 
> require CC1 and CC2. I am thinking of adding AB block to the protocols. Do 
> you have any other suggestions?. I use the ULTRA stainers.
> 
> Thank you very much in advance,
> 
> Milton A. Gomez, HTL (ASCP)
> Technical Supervisor
> Immunohistochemistry Department
> ARUP Laboratories, Inc.
> 500 Chipeta Way
> Salt Lake City, UT 84108-1221
> Desk Phone: 801-583-2787, ext.3869
> Lab. Phone: 801-584-5257/5242
> Fax: 801-584-5217
> E-mail: milton.go...@aruplab.com  
> Web: www.aruplab.com  
> 
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> attachments are from ARUP Laboratories and are intended only for the
> recipient. The information contained in this message is confidential
> and may constitute inside or non-public information under
> international, federal, or state securities laws, or protected health
> information and is intended only for the use of the recipient.
> Unauthorized forwarding, printing, copying, distributing, or use of
> such information is strictly prohibited and may be unlawful. If you
> are not the intended recipient, please promptly delete this e-mail
> and notify the sender of the delivery error or you may call ARUP
> Laboratories Compliance Hot Line in Salt Lake City, Utah USA at (+1
> (800) 522-2787 ext. 2100
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RE: [Histonet] FNA stain

[Tony Henwood]

We use the Diff quik (or whatever spelling you use!)stain on air-dried
smears.

There is also a rapid Poor man's H&E stain, using the Diff Quik
solutions, that you can also use. You can then restain with the normal
PAP stain back in the lab (Hirschowitz et al Acta Cytolog (1994)
38:499-501)

We try to give an immediate provisional diagnosis at the time

Regards

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




-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
dkb...@chs.net
Sent: Thursday, 17 December 2009 5:24 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] FNA stain


This question is for those of you who perform fine needle aspirations. 
What stain are you using for your immediate evaluation?  Or do you give
an 
immediate evaluation/adequacy?
Thanks.

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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[Histonet] looking for KOH nail procedure

[Cheryl]

 Hi Guys!
 
I've used a potassium hydroxide solution to 'soften' nails after fixation but 
before processing.  I cannot find my procedure!!
 
Can anyone help?
 
Cheryl Kerry, HT(ASCP)
 
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[Histonet] IMAGING

[oscar gonzalez]

If you have seen the new CAP inspection lists, there are several questions in 
regards to Imaging. Does anybody have a procedure to cover such questions?
Thanks.


  
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[Histonet] immunoperoxidase background staining

[Gomez, Milton]

Dear Histonetters,
 
I am getting background staining with my Immunohistochemistry protocols that 
require CC1 and CC2.  I am thinking of adding AB block to the protocols.  Do 
you have any other suggestions?.  I use the ULTRA stainers.
 
Thank you very much in advance,
 
Milton A. Gomez, HTL (ASCP)
Technical Supervisor
Immunohistochemistry Department
ARUP Laboratories, Inc.
500 Chipeta Way
Salt Lake City, UT 84108-1221
Desk Phone:  801-583-2787, ext.3869
Lab. Phone:   801-584-5257/5242
Fax:  801-584-5217
E-mail:  milton.go...@aruplab.com  
Web:  www.aruplab.com  

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The information transmitted by this e-mail and any included
attachments are from ARUP Laboratories and are intended only for the
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and may constitute inside or non-public information under
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information and is intended only for the use of the recipient.
Unauthorized forwarding, printing, copying, distributing, or use of
such information is strictly prohibited and may be unlawful. If you
are not the intended recipient, please promptly delete this e-mail
and notify the sender of the delivery error or you may call ARUP
Laboratories Compliance Hot Line in Salt Lake City, Utah USA at (+1
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Re: [Histonet] leaving IHC slides in wash buffer

[Rene J Buesa]

I would never do that. Why subject your sections to such a treatment?
René J.





From: Jennifer Campbell 
To: histonet@lists.utsouthwestern.edu
Sent: Wed, December 16, 2009 1:34:40 PM
Subject: [Histonet] leaving IHC slides in wash buffer

Hi Everyone,

  I have yet another IHC-related question.  So after antigen retrieval,
and then allowing my slides too cool, I place them in wash buffer before
loading them on to the autostainer.  Would leaving the slides in the
wash buffer in the fridge for an extended period of time (say, 1 week)
before continuing with immunostaining affect staining quality in any
way?

Thank you,

Jen Campbell
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[Histonet] RE: A good "lean" consultant for a pathology lab

[Mahoney,Janice A]

I'd highly recommend Randy Stephens.
randysteph...@mac.com

Jan Mahoney, Omaha, NE

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Perkocha, Luke
Sent: Wednesday, December 16, 2009 12:57 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: [Histonet] A good "lean" consultant for a pathology lab

We are a multi-site academic AP-only laboratory and are looking for 
recommendations for process improvement consultants/trainers with experience 
with Lean, Six-sigma and other process improvement tools. We would like to 
bring someone in initially to help with a couple of small projects, with goals 
of:

 1.  Improving both process and physical environment to increase efficiency and 
reduce the chance of error (improve patient safety) in the EM lab.
 2.  Simultaneously providing some training, so that some tools and 
competencies are left over in the Department in this area.

If this is a good experience, we would like to look at larger divisions (i.e. 
gross room, histology, specimen transport and receipt, professional sign-out, 
etc.) or more of the overall production process.

There are so many consultants out there. Can anyone recommend someone or a 
group that is both competent and cost-effective, and can handle both the 
project itself, plus some training around it - hopefully with a track record of 
success in similar lab situations? Many thanks for your help!

Luke Perkocha
luke.perko...@ucsf.edu
415 885-7254
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[Histonet] FW: FW: Follow Up: Joint Commission Lab Survey CorrectiveAction Plan West MOHS

[Ingles Claire]

OK Guys, here's another JCAHO/CLIA controversy. Has anyone (Mostly MOHS labs) 
been sited for this? Currently we only save tissue specimens 24 hours in case 
the Dr. decides to send some to path after the fact. (don't get me going on 
morphology quality). Does anyone have a clear definition of "gross tissue 
specimens" It's not like we measure this stuff before freezing.
Thanks for your well educated guesses
Claire


 

West MOHS Clinic was cited for a deficiency regarding maintenance and storage 
of tissue during the Joint Commission Lab Survey. 

According to standard QC.2.120, EP 2:  Microscopic slides, paraffin blocks, 
bone marrow aspirates, needle biopsy specimens, and gross tissue specimens are 
stored for required times as defined by organization policy, law, and 
regulation.  Microscopic slides, including stained slides, are retained for at 
least 10 years.  Paraffin blocks are stored for a minimum of two years from the 
date of examination.  Gross tissue specimens are retained for at least seven 
days after required microscopic sections are examined and reports reviewed and 
signed.  State law and regulation requirements are for longer times (See 
Appendix XX for additional specific retention times.)

The surveyor observed in his report that "biopsy tissue was frozen for section, 
slides prepared and then the tissue was discarded.  Gross tissue specimens must 
be retained for seven days after the microscopic sections are examined and 
reports reviewed and signed."


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[Histonet] Re: COX2 in mice

[Hobbs, Carl]

What are your experimental conditions?
carl



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[Histonet] VZV Controls

[Jack Dodo]

Does anyone have VZV Controls? A block would be great, but at this point I will 
take anything. Can anyone point me in the right direction in finding this? 
Thanks 
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[Histonet] A good "lean" consultant for a pathology lab

[Perkocha, Luke]

We are a multi-site academic AP-only laboratory and are looking for 
recommendations for process improvement consultants/trainers with experience 
with Lean, Six-sigma and other process improvement tools. We would like to 
bring someone in initially to help with a couple of small projects, with goals 
of:

 1.  Improving both process and physical environment to increase efficiency and 
reduce the chance of error (improve patient safety) in the EM lab.
 2.  Simultaneously providing some training, so that some tools and 
competencies are left over in the Department in this area.

If this is a good experience, we would like to look at larger divisions (i.e. 
gross room, histology, specimen transport and receipt, professional sign-out, 
etc.) or more of the overall production process.

There are so many consultants out there. Can anyone recommend someone or a 
group that is both competent and cost-effective, and can handle both the 
project itself, plus some training around it - hopefully with a track record of 
success in similar lab situations? Many thanks for your help!

Luke Perkocha
luke.perko...@ucsf.edu
415 885-7254
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[Histonet] good Calcafluor & Auramine/Rhodamine staining method for FFPE tissue

[Sharon Allen]

Hi, I am posting this question for Lisa,

Our Microbiology department would like to find a good calcafluor
staining method and a good Auramine/Rhodamine staining method for FFPE
tissues.  Any suggestions?  Would you be able to post this for me?
  Thanks, Lisa

Lisa Manning MLT, BSc.
Pathology Technical Director
Diagnostic Services of Manitoba
401B Brodie Centre
727 McDermot Avenue
Winnipeg, Manitoba
R3E 3P5
Ph. (204) 789-3325
Fax (204) 789-3931
lmann...@hsc.mb.ca

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[Histonet] restaining IHC slides

[Thomas Pier]

Hello Histonetters,
I ran into an issue when staining some TMA slides the other day. This question 
isn't about how to fix the staining, or lack there of. I would like to know 
whether or not it is possible to repeat the same protocol on the same slides 
after removing the coverslips. If any of you have any experience with this sort 
of thing, please let me know.

Tom Pier
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Re: [Histonet] leaving IHC slides in wash buffer

[V. Neubert]

I guess yes.

Unless I can say exactly which ingredient will affect my staining, I 
would not leave them in any solution for such a long time!



Hi Everyone,

   I have yet another IHC-related question.  So after antigen retrieval,
and then allowing my slides too cool, I place them in wash buffer before
loading them on to the autostainer.  Would leaving the slides in the
wash buffer in the fridge for an extended period of time (say, 1 week)
before continuing with immunostaining affect staining quality in any
way?

Thank you,

Jen Campbell
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[Histonet] leaving IHC slides in wash buffer

[Jennifer Campbell]

Hi Everyone,
 
  I have yet another IHC-related question.  So after antigen retrieval,
and then allowing my slides too cool, I place them in wash buffer before
loading them on to the autostainer.  Would leaving the slides in the
wash buffer in the fridge for an extended period of time (say, 1 week)
before continuing with immunostaining affect staining quality in any
way?
 
Thank you,
 
Jen Campbell
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[Histonet] Repairs to equipment.

[Oeler, Theresa]

 
Hi,  We use Tech One Biomedical Services, Inc. as a third party PM and
repair service. They are very nice to work with and not excessive with
their fees. We have set up PM's with them for a lot of our older
equipment. Toll free number is 866-497-3033 or www.techoneweb.com . Try
them for your repairs, we have no problems with them for the four years
we have been dealing with them.  

No trees were harmed in the sending of this email. However, many
electrons were severely inconvenienced.
Theresa A Oeler, BS
Senior Research Histologist
NSABP Pathology Laboratory
Federal North Building
1307 Federal Street, Suite 303
Pittsburgh, PA 15212
412/359-8931 Of.  412/359-3239 Fx.
theresa.oe...@nsabp.org

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
histonet-requ...@lists.utsouthwestern.edu
Sent: Wednesday, December 16, 2009 1:20 PM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 73, Issue 24

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To subscribe or unsubscribe via the World Wide Web, visit
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When replying, please edit your Subject line so it is more specific than
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Today's Topics:

   1. FW: Notch-Nodal Paper (Margaryan, Naira)
   2. RE: Blade Dispenser Thingy (Bonner, Janet)
   3. RE: Blade Dispenser Thingy (Maria Katleba)
   4. Histology Jobs Outside the USA (Maria Katleba)
   5. COX2 in mice (Bell, Pat)
   6. Anti-Flag antibody (Rachel, Rivka (NIH/NEI) [E])
   7. Leica Bond (Liz Chlipala)
   8. RE: Leica Bond (Debrosse-Serra, Beatrice)
   9. Re: FW: Notch-Nodal Paper (John Kiernan)
  10. any info on Glutathione S-Transferase Pi (GST Pi)?
  (Jennifer Campbell)
  11. Equipment Repair (Yaskovich, Ruth A (NIH/NIDCR) [E])


--

Message: 1
Date: Tue, 15 Dec 2009 12:00:54 -0600
From: "Margaryan, Naira" 
Subject: [Histonet] FW: Notch-Nodal Paper
To: "histonet@lists.utsouthwestern.edu"

Message-ID:



Content-Type: text/plain; charset="us-ascii"

Hi histonetters,

I am looking for some dye to be used in vivo like cell tracker or
similar might work for identifying live cells.

Any suggestions are appreciated,
Naira


--

Message: 2
Date: Tue, 15 Dec 2009 13:51:23 -0500
From: "Bonner, Janet" 
Subject: RE: [Histonet] Blade Dispenser Thingy
To: "Breeden, Sara" ,
histonet@lists.utsouthwestern.edu
Message-ID:

<5f31f38c96781a4fbe3196ebc22d47807f2...@fhosxchmb006.adventistcorp.net>

Content-Type: text/plain; charset=iso-8859-1

I keep the dispenser in a small plastic bag that seals along with the
desiccant packages they put in the original box with the blades, (and
any other desiccant packages I can find - from coverslips, inks etc.)
As long as you keep the blades sealed with the desiccant, you shouldn't
have anymore problems.  This works really well in the cryostat chamber,
too!
 
Janet 



From: histonet-boun...@lists.utsouthwestern.edu on behalf of Breeden,
Sara
Sent: Tue 12/15/2009 9:52 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Blade Dispenser Thingy



I have Feather dispensers with blades that I have to pry out, which is
probably not a good thing.  I've tried freezing the dispenser, putting
it in an oven for a while and whacking the darned thing on the floor
(not necessarily all at the same time..).  Have we had this discussion
before and have we solved this problem?  Help?



Sally Breeden, HT(ASCP)

NM Dept. of Agriculture

Veterinary Diagnostic Services

PO Box 4700

Albuquerque, NM  87106

505-841-2576



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confidential and protected from disclosure.  If the reader of this
message is not the intended recipient or an employee or agent
responsible for delivering this message to the intended recipient, you
are hereby notified that any dissemination, distribution or copying of
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Message: 3
Date: Tue, 15 Dec 2009 11:39:48 -0800
From: Maria Katleba 
Subject: [Histonet] RE: Blade Dispenser Thingy
To: "Bonner,

[Histonet] FNA stain

[DKBoyd]

This question is for those of you who perform fine needle aspirations. 
What stain are you using for your immediate evaluation?  Or do you give an 
immediate evaluation/adequacy?
Thanks.

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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[Histonet] New processor help

[Carole Ruffin]

Is there a procedure or some standard method validation process out there
for starting up a new tissue processor? Maybe one that would be approved
by regulatory agency? 
I am not sure whether this would just be left up to the pathologists to 
decide what they would prefer.

Thank you,

Carole Ruffin, HT(ASCP)
ValleyCare Hospital
Pleasanton, CA







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[Histonet] Equipment Repair

[Yaskovich, Ruth A (NIH/NIDCR) [E]]

Does anyone know of a company that repairs equipment in the Maryland area? I'm 
in Bethesda and my old Tissue-Tek embedding center won't hold the temperature.
Thanks for any help,
Ruth Yaskovich
National Institutes of Health
Bethesda, Maryland
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[Histonet] any info on Glutathione S-Transferase Pi (GST Pi)?

[Jennifer Campbell]

Hi All,
 
  Has anyone ever used this antibody in FFPE mouse or rat tissue?  How
easy of an antibody is it to work with?  Other comments?  We may be
doing this for an upcoming project and I just wanted to know what I
could be getting myself into.  Thanks!
 
Jen Campbell
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