Re: [Histonet] decal protocol with EDTA for bone marrow biopsies

[Пешков Максим via Histonet]

We fixed our BM biopsies at least 24 hours, but no more than 72 h in 10% NBF.
Wash with tap water 30 mins.
Decal into 14% EDTA (2Na salt) buffered with NaOH up to 7.0 overnight or 
weekend and test endpoint by weight loss weight gain method (special thanks to 
Gayle Callis!).
Wash in tap water 1 hour.
Process with isopropanols (IPA), mixtures IPA and mineral oil), paraffin wax.
Microtomy at 3 microns.
Stain HE, Giemsa, HC, IHC.
--
Maxim Peshkov,
Russia,
Taganrog.
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[Histonet] Quality Measures and Productivity Trackers for Anatomic Pathology Program Support Assistants

[Dachel, Susan K. via Histonet]

Does anyone have any quality measures or productivity trackers that they have 
used for Anatomic Pathology office staff?  I am particularity interested in  
error rate acceptability and productivity standards that include typing gross 
and microscopic dictation, entering SNOMED and CPT coding, and entering 
supplemental or modified reports.

Thanks in advance for your help!
Sue

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[Histonet] FW: Histonet post

[Linda Margraf via Histonet]

Here is a message I am posting for Samantha. Please reply to her not me, thanks!
From: "Green, Samantha" 
>
Date: August 28, 2017 at 3:18:32 PM CDT
To: 'Linda Margraf' >
Subject: Histonet post
Hello Histoneters,

I am reaching out to the pathology world in the hopes that someone can help us 
out. Our lab is currently validating two new staining protocols:


1.   HER2 FISH for gastric tissue (stomach & distal esophagus)

2.   ALK FISH for lung tissue

We have hit a road block without validation because we have run out of patient 
tissue. We are desperately searching for:


· 10 distal esophagus / gastric tissue POSITIVE for HER2 (preferably 
stained by FISH previously)

· 5 distal esophagus / gastric tissue NEGATIVE for HER2 (preferably 
stained by FISH previously)

· 20 specimens (surgical or cytology cell block) POSITIVE for ALK 
(previously stained by FISH only)
We would need two charged slides with 5 micron sections and proof of stain 
results (example: de-identified report).

We understand this is a lot of work but anything thing would help, even if it 
is just one or two cases. We are not asking anyone to “work for free” so we are 
100% willing to trade help for tissue blocks or purchase the slides if 
necessary. We do have TONS of FFPE tissue blocks that are HSV I & II positive. 
I know that HSV can be hard to come by and we would be willing to send some 
tissue blocks in exchange for the slides listed above.

Please either post on this blog or contact me directly at 
samantha.gr...@cmc-nh.org .

Thanks for reading!

Best regards -

Samantha Green, B.S.  HTL (ASCP)


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Re: [Histonet] Paraformaldehyde Fixed Tissue

[Michael Gudo via Histonet]

Dear Sandra,
Dear Ana,

in our lab for tissues for which we have enough time for infiltration we fix 
for at least 48 hours or more in NBF or PFA (both with 4 % reactive 
formaldehyde, which is equivalent to 10% „neutral buffered formaline“) and then 
first wash in tap water for a minimum of 1-2 hours, then to 30%, 50%, 60%, 70%, 
80%, 90% 96% (2 times), 2-Propanol (2-times) and then 2-3 time xylene, before 
embedding in paraffine. The times are just a few hours per step or shorter, and 
we have quite good results especially for brain and spinal chord.

Best regards
Michael



> Am 29.08.2017 um 02:06 schrieb Ana Maluenda via Histonet 
> :
> 
> Hi Sandra,
> 
> I haven't myself particularly worked with brain and spinal cord, but majority 
> of my protocols in my old job used fixation in 4% PFA (24 hours at 4-8oC) and 
> routinely process (or transfer to graded EtOH, if not processing immediately).
> However, our routine process didn't include a first step in 10% NBF, since 
> PFA plays the role of fixation. Therefore, after PFA, we would have 70% EtOH, 
> 80% EtOH, 95% ETOH, 2x EtOH the Xylenes and wax [assuming you are referring 
> to FFPE?].
> Also mouse tissue can be small and delicate, so I remember running liver, 
> spleen, kidney and thymus (soft tissues I worked with) in a short cycle 
> (similar to what I would do for biopsies).
> 
> Hope this helps!
> 
> Kind regards,
> 
> Ana
> 
> 
> Ana Maluenda
> Research Assistant
> Atherothrombosis and Vascular Biology Laboratory
> 
> Baker Heart and Diabetes Institute
> 75 Commercial Road, Melbourne VIC 3004
> P (03) 8532 1359 E ana.malue...@baker.edu.au W www.baker.edu.au
> 
> -Original Message-
> From: Sandra Cheasty [mailto:sandra.chea...@wisc.edu]
> Sent: Tuesday, 29 August 2017 1:30 AM
> To: Histonet (histonet@lists.utsouthwestern.edu) 
> 
> Subject: [Histonet] Paraformaldehyde Fixed Tissue
> 
> Hi all,
>   We are having difficulty sectioning mouse tissue, (brain, 
> spinal cord, liver, and spleen), on paraformaldehyde fixed tissue. Has anyone 
> had issues with paraformaldehyde fixed tissue? They were processed routinely, 
> starting in 10% NBF, with other tissues, and we are cutting them at 3u.
> Thank you!
> Sandy
> 
> Sandra J. Cheasty, HT (ASCP)
> Histology & Necropsy Supervisor
> UW-Madison, School of Veterinary Medicine
> 
> 
> Protecting your privacy is important to us. The Baker Heart and Diabetes 
> Institute will handle your information in accordance with the Privacy Act 
> 1988 (Cth) and its Privacy Policy which is available at www.baker.edu.au or 
> on request by contacting priv...@baker.edu.au or by calling 1800 838 498. The 
> Privacy Policy also explains how you can access and correct your personal 
> information, or make a complaint about a breach of the Australian Privacy 
> Principles. bidipp2014.0.1a
> -- 
> Message  protected by MailGuard: e-mail anti-virus, anti-spam and content 
> filtering.http://www.mailguard.com.au/mg
> 
> 
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Re: [Histonet] Cut FFPE Slides for ISH

[Michael Gudo via Histonet]

Hello Jenniffer,

we store the slides with section at 4-8 °C for many month or even longer and we 
do not have any problems with ISH.

Best regards
Michael



> Am 28.08.2017 um 17:30 schrieb Jennifer Phinney via Histonet 
> :
> 
> Hello Histonet,
> I am hoping someone can point me in the right direction to find some 
> references for how long FFPE slides intended for ISH can be kept before 
> staining.  I have a researcher who swears their slides have to be stained 
> within 24 hours of being cut, which does not sound correct to me.  I've used 
> ACD's RNAscope before and their protocol lists the timeframe as 3 months (so 
> this is a bit of a difference).
> 
> There's a lot of information about fixation times, but I've come up empty 
> handed for cut slides.
> 
> Thanks everyone!
> 
> Jennifer Phinney QIHCCM
> Project Coordinator
> Kansas State University
> Veterinary Diagnostic Lab
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[Histonet] decal protocol with EDTA for bonemarrow biopsies

[Gudrun Lang via Histonet]

Dear histonetters!

I would be happy about some input about decalcification protocols with EDTA
of trephine bonemarrow biopsies.

recommended duration of fixation?

recommended duration of decalcification?

strategies for speed-up of decal?

recommended EDTA-solution formula?

 

Hopefully some experienced histotechs can share their knowledge with me.

thanks in advance

Gudrun

 

 

 

 

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[Histonet] CAP checklist question

[Vanessa Keeton via Histonet]

Good morning fellow histonetters!

I was wondering if anyone in VA would know the laws in reference to
checklist question ANP. 10016 in regards to the exclusion of submission of
tissue and removal of objects for Pathology examination?

CAP gave an example where California Department of Health Care Services
requires all tissues and objects removed during surgery to be submitted for
Pathology examination, unless a specific request is submitted to the state
requestinga variance.

Any insight would be greatly appreciated.

Vanessa Keeton
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