[Histonet] Histonet Digest, Vol 147, Issue 31

k.leigh.adams.865 k.leigh.adams.865 at gmail.com
Mon Feb 29 16:14:01 CST 2016


    
Ted Pella, pelco freeze spray.



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-------- Original message --------
From: histonet-request at lists.utsouthwestern.edu 
Date: 2/29/2016  1:00 PM  (GMT-05:00) 
To: histonet at lists.utsouthwestern.edu 
Subject: Histonet Digest, Vol 147, Issue 31 

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Today's Topics:

   1. H&E staining (Charles Riley)
   2. Job openning in Chicago Suburbs (Lester Raff MD)
   3. H&E staining (Carlos Defeo)
   4. Re: H&E staining (Rene J Buesa)
   5. NJ Histology Meeting in June (FRENCH, MICHELE)
   6. Freeze Spray Ban (Histology)


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Message: 1
Date: Mon, 29 Feb 2016 06:15:54 -0500
From: Charles Riley <criley at dpspa.com>
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] H&E staining
Message-ID:
	<CAAQhB13iuzFq0XQFrU9cDVeLp+42M1aTiX13t8Ao4JtZ5-FwCA at mail.gmail.com>
Content-Type: text/plain; charset=UTF-8

Hello all,

 We've been running into an issue lately with our H&E staining. The
hematoxylin and eosin are light on and off but the chromatin is
consistently staining light. I have changed the time in hematoxylin to
longer, used a fresh lot of both hematoxylin and our clarifier solution and
the stain is not improving.  Does anyone have any suggestions?

Here is our routine stain line procedure.
65 degree Celsius oven for 20 minutes
2x xylene for 5 mins
2x 100% alcohol for  1 minute
95% alcohol for 1 minute
DI water rinse for 1 minute
Richard Allan scientific hematoxylin for 4 minutes
DI water rinse for 2 minutes
Richard Allan Clarifier 1 for 2.5 minutes
DI water for 2 minutes
Bluing Reagent for 1 minute
DI water rinse for 2 minutes
95% alcohol for 1 minute
Eosin Y for 2 minutes
95% alcohol for 30 seconds
2x 100% alcohol for 1 minute each
2x xylene for 1 minute each
Mount and coverslip.
-- 

Charles Riley HT(ASCP)CM

Histopathology Coordinator/ Mohs

Doctors Pathology Services, Dover DE


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

Message: 2
Date: Mon, 29 Feb 2016 13:12:19 +0000
From: Lester Raff MD <LRaff at uropartners.com>
To: "'histonet at lists.utsouthwestern.edu'"
	<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Job openning in Chicago Suburbs
Message-ID:
	<6347C6D2B080534F9B5C2B08436DCFAF0B705FC0 at COLOEXCH01.uropartners.local>
	
Content-Type: text/plain; charset="us-ascii"

Our lab, a large private uropathology lab in the west Chicago suburbs, is seeking a full time day shift histologist.  Please contact me for details. Please spread the word!


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: 3
Date: Mon, 29 Feb 2016 13:31:35 +0000
From: "Carlos Defeo" <latecor at adinet.com.uy>
To: "Charles Riley via Histonet" <histonet at lists.utsouthwestern.edu>
Cc: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] H&E staining
Message-ID: <em57c9e61c-45a7-4e63-b4e0-7a5e2af5384d at calipc>
Content-Type: text/plain; format=flowed; charset=utf-8

Charles,extend time on the deparaffinization station,and even 
better,renew solvents at this point also.
My kind regards,
Carlos Defeo
Histotechnologist

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

Message: 4
Date: Mon, 29 Feb 2016 14:27:25 +0000 (UTC)
From: Rene J Buesa <rjbuesa at yahoo.com>
To: Charles Riley <criley at dpspa.com>,
	"histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] H&E staining
Message-ID:
	<1567788252.777382.1456756045073.JavaMail.yahoo at mail.yahoo.com>
Content-Type: text/plain; charset=UTF-8

Usually inconsistency in H&E staining has two different sources: either the staining process itself, or the previous tissue processing sequence.Your staining protocol has a "standard" sequence but I would add an additional xylene and would increase the time in hemtoxylin to 7 minutes for you will differentiate ant way with the clarifier. Are you staining manually? If so the time in the clarifier should not be standard, the stained sections should remain in it until no more color leaches from the sections and should be as short as possible. Most likely your staining problems resides in the "clarifier" step and when the sections are of the "right" tissue type and the "right" thickness the staining is OK, but when they are either too thin or of tissues with less amounts of nuclei, the staining is faint.
In any even, I strongly suggest you to change your whole protocol and eliminate xylene and alcohols in the dewaxing step, and switch to a 2% aq. sol. of dishwashing soap at 90?C (2 stations x 2 min. each) ? hot distilled water (90?C x 1 ? 45?C x 1 min) ? dist. water at room temp. which will allow you not only eliminating xylene but also ?finish the whole procedure sooner and much cheaper.Try it!

Ren? 

    On Monday, February 29, 2016 6:33 AM, Charles Riley via Histonet <histonet at lists.utsouthwestern.edu> wrote:
 

 Hello all,

 We've been running into an issue lately with our H&E staining. The
hematoxylin and eosin are light on and off but the chromatin is
consistently staining light. I have changed the time in hematoxylin to
longer, used a fresh lot of both hematoxylin and our clarifier solution and
the stain is not improving.? Does anyone have any suggestions?

Here is our routine stain line procedure.
65 degree Celsius oven for 20 minutes
2x xylene for 5 mins
2x 100% alcohol for? 1 minute
95% alcohol for 1 minute
DI water rinse for 1 minute
Richard Allan scientific hematoxylin for 4 minutes
DI water rinse for 2 minutes
Richard Allan Clarifier 1 for 2.5 minutes
DI water for 2 minutes
Bluing Reagent for 1 minute
DI water rinse for 2 minutes
95% alcohol for 1 minute
Eosin Y for 2 minutes
95% alcohol for 30 seconds
2x 100% alcohol for 1 minute each
2x xylene for 1 minute each
Mount and coverslip.
-- 

Charles Riley HT(ASCP)CM

Histopathology Coordinator/ Mohs

Doctors Pathology Services, Dover DE
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


  

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

Message: 5
Date: Mon, 29 Feb 2016 17:12:37 +0000
From: "FRENCH, MICHELE" <MICHELE.FRENCH at bms.com>
To: "Histonet (histonet at lists.utsouthwestern.edu)"
	<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] NJ Histology Meeting in June
Message-ID:
	<0cf99f07da5f48cfa117b4dfe4ab8b85 at DM2PR26MB0030.067d.mgd.msft.net>
Content-Type: text/plain; charset="us-ascii"

Dear Histonet Members,

I wanted to let you know that The New Jersey Society for Histotechnology is planning to host a two-day Summer Meeting June 10-11 at the Wyndham Hotel in Mount Laurel, NJ. More information will be available at the end of March, and meeting registration will start on April 1st.

SPEAKERS NEEDED ASAP: We need to fill out last three time slots: (one 1.5hr session, one 3hr session and one 45minute session).
VENDORS NEEDED: We only have 14 tables left in the exhibit area. Reservations are on a first-come, first-served basis.
JOB LISTINGS WANTED: If you have a job opening that you would like posted on our Employment Opportunities board during the event, just let us know!

Please contact Michele French by Email Michele.French at bms.com<mailto:Michele.French at bms.com> or Phone 609-818-3873.

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

Message: 6
Date: Mon, 29 Feb 2016 17:29:34 +0000
From: Histology <histo at pathlab.us>
To: "histonet at lists.utsouthwestern.edu"
	<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Freeze Spray Ban
Message-ID:
	<09CFA3F99D5B2B42B88CDFB2FC4CFD823433DD at vdc01.domain.local>
Content-Type: text/plain; charset="us-ascii"

Hi All,

Just got an alert stating that the EPA is banning HFCs found in Freeze Spray effective July 20th.  Does anybody use a spray that does not contain this banned chemical?  If so, please let me know where you get it.

Thanks,

Mehndi Helgren
Dominion Pathology Labs  "DPL"


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