[Histonet] CD34 for primate tissue

Fri Jul 1 14:16:32 CDT 2016

This is a great website for NHP info.  Thank you all for all your wonderful help!!
Hope everyone has a great 4th of July weekend!!!!
Cathy

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To: histonet at lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 152, Issue 1

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Today's Topics:

   1. Re: How to get your stains more vibrant when cutting at 2?m?
      (Angela Lamberth)
   2. Re: CD34 for primate tissue (Croix, Denise)
   3. Just some before the long weekend Friday Fun-lab related	blog
      (Lester Raff MD)
   4. Certification (Adesupo, Adesuyi (Banjo))

----------------------------------------------------------------------

Message: 1
Date: Thu, 30 Jun 2016 10:41:38 -0700
From: Angela Lamberth <alamberth at lji.org>
Cc: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] How to get your stains more vibrant when
	cutting at 2?m?
Message-ID:
	<CANUNjWni6rSWkGqvp5u7-WN3R29Gk4mOV_gr-YkWa41vtjt1GA at mail.gmail.com>
Content-Type: text/plain; charset=UTF-8

Thank you all very much for your suggestions. I'm going to play around with a progressive H&E when I return from vacation next month. I do have safranin on hand but will need to order some phloxine to experiment with. I will probably need to order some additional supplies to make the hematoxylin. I'm not sure yet if I'll start with Mayer's or Gill's or maybe even Erlich's.

I should note that cutting at 2 isn't required, but it is desired once they saw that I can. And since I can, I aim to please! :-)

In addition, I'm looking forward to trying the oven dry method before coverslipping. A rapid dehydration isn't really possible since I'm working with a xylene substitute (Pro-Par) and have been battling eosin carryover but that is a whole different animal for another thread.

Thanks again! I'll report back in a month or 2.

On Wed, Jun 29, 2016 at 10:27 AM, Rene J Buesa <rjbuesa at yahoo.com> wrote:

>
> Angela:
> "Pale" results are the trade-off for great quality very thin "2 ?m"
> sections but you can always improve intensity somewhat .
> 1- your "regressive" stain, if it is "modern Harris" has the inherent 
> problem of lacking mercury chloride and it is little you can do about.
> Perhaps if you use "progressive Mayer" you will get better results. 
> You will not have to differentiate (with the intrinsic "danger" of 
> leaving the section too pale) and if used fresh Mayer's can be a good approach.
> 2- as to the counterstain perhaps you should add safranine to the 
> eosin (20% safranine + 80% eosin) and will get a darker red.
> 3- try to dehydrate as quickly as possible or even better, wash the 
> sections in distilled water and place them in an oven at 60?C for 10 
> minutes and coverslip as usual. You will eliminate any "color wash" 
> due to the alcohols.
> If you've not enough "trust" on dry/oven dehydration, try with some 
> sections as a test. You will like the method.
> Ren?
>
>
> On Tuesday, June 28, 2016 5:53 PM, Angela Lamberth via Histonet < 
> histonet at lists.utsouthwestern.edu> wrote:
>
>
> When I cut at 2?m my H&Es and special stains look pale. How can I get 
> my stains to pop or am I stuck with pale looking stains when 
> sectioning that thin?
>
> I run manual specials and a manual regressive H&E. For H&E I've tried 
> increasing my time in hematoxylin (beyond the manufacturer 
> recommendation), diluting my acid alcohol differentiation, and 
> increased time in eosin but the slides still lack the vibrancy that many of the postdocs desire.
>
> I use Shandon instant hematoxylin and alcoholic eosin by Thermo. 
> Everything else I prepare in house from scratch. Any recommendations?
> _______________________________________________
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> Histonet at lists.utsouthwestern.edu
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>
>

--
Angela Lamberth
Histology Technician II
Histology Core Lab
La Jolla Institute for Allergy & Immunology
9420 Athena Circle
La Jolla, CA 92037

------------------------------

Message: 2
Date: Thu, 30 Jun 2016 16:57:53 -0500
From: "Croix, Denise" <denise.croix at roche.com>
To: histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] CD34 for primate tissue
Message-ID:
	<CABifpndB_vHDwpZDYfm9_5xOmNOZf9ktwXDpod+qiQ5okva3rQ at mail.gmail.com>
Content-Type: text/plain; charset=UTF-8

Hi Cathy,

When I was doing work with NHP tissue (both flow cytometry and IHC) there
were several sources that I utilized for info on antibodies that were
cross-reactivity with various NHP species.  The NIH hosts a NHP reagent
resource website (nhpreagents.org) that shows which commercially available
antibodies cross react with non-human primate tissues.  They note that
there are discrepancies reported about cross-reactivity with the QBEND10
antibody for  rhesus and possibly cynos.  You will need to still check with
the vendors to confirm if the antibody has been tested in FFPE specimens as
that is not indicated on the site but it should give you a starting point
to work from.

I've had personal experience with cyno & rhesus T cells where individual
monkeys would not react with certain anti-human antibodies.  For example, I
had a couple of research animals that were negative for a CD3 antibody but
yet they stained with CD4 and CD8 antibodies.

Good luck!

Denise

-- 
[image: Powered by Sigstr]
<https://rochediagnostic.sigstr.net/uc/56dd9d91825be9040100000b/watermark>*Denise
A Croix, Ph.D.*
Pathology Solutions Specialist - PD-L1 SP142 Assay
Roche Diagnostics Corporation
Tissue Diagnostics Division

972-207-7305
denise.croix at roche.com
www.ventana.com

[image: Powered by Sigstr]
<https://rochediagnostic.sigstr.net/uc/56dd9d91825be9040100000b/watermark>
<https://rochediagnostic.sigstr.net/uc/56dd9d91825be90401000032>
[image: Powered by Sigstr]
<https://rochediagnostic.sigstr.net/uc/56dd9d91825be90401000032/watermark>

------------------------------

Message: 3
Date: Fri, 1 Jul 2016 13:44:10 +0000
From: Lester Raff MD <LRaff at uropartners.com>
To: "Histonet at lists.utsouthwestern.edu"
	<Histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Just some before the long weekend Friday Fun-lab
	related	blog
Message-ID:
	<6347C6D2B080534F9B5C2B08436DCFAF10E5E6FC at COLOEXCH01.uropartners.local>

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

Pathologists couldn't do it without great histology teams, so thanks and have a Happy 4th.

http://www.chicagonow.com/downsize-maybe/2016/06/reading-a-prostate-biopsy-inside-the-mind-of-a-pathologist/

Lester J. Raff, MD MBA
UroPartners
Medical Director Of Laboratory
2225 Enterprise Dr. Suite 2511
Westchester, Il 60154
Tel: 708-486-0076
Fax: 708-492-0203

------------------------------

Message: 4
Date: Fri, 1 Jul 2016 09:45:31 -0500
From: "Adesupo, Adesuyi (Banjo)" <abadesuyi at nrh-ok.com>
To: "'histonet at lists.utsouthwestern.edu'"
	<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Certification
Message-ID:
	<04EE4F75BB5FB246ADB68D69B7460443AF7A2CEA42 at MAIL.nrhnt.nrh-ok.com>
Content-Type: text/plain; charset="us-ascii"

Hi,
    I hope you guys are doing great. Please I have a question and would appreciate your contributions. I have a tech that could not pass the certification test, when the ASCP were still accepting high school diploma for HT.
The tech is no longer eligible to take the test again, because he did not have the minimum requirement (i.e. Associate Degree) to register for the test. He wants me to promote him to Histology Lead Tech/Histology Coordinator.
What do you guys think about this?

Best regards,

Banjo Adesuyi, BMLS, HT (ASCP) HTL, QIHC, QLS

Histology Supervisor

Norman Regional Health System,

Norman, OK 73071.

Tel: 405- 307- 1145

Cell: 405-973-6363

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