[Histonet] RE: H&E on Frozen

[Rathborne, Toni]

Our protocol is to:

Use a positively charged slide
Fix in 10% formalin for 1 minute
Rinse in tap (a few dips)
Gill 3 for 1 minute
Rinse in tap (a few dips)
3-5 dips in bluing
Rinse in tap (a few dips)
Eosin 2-3 dips
100% ETOH (a few dips)
100% ETOH (a few dips)
Xylene (dip until clear)
Mount with Richard- Allan Mounting Medium

After the Gill3, Eosin, and last alcohol, the back of the slide is wiped clean of excess solution to prevent carryover.


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Reynolds,Donna M
Sent: Tuesday, August 16, 2011 10:13 AM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] H&E on Frozen

We do this frequently. 
Fix slides in Methanol (45ml) + formaldehyde (5ml) for 10 minutes. 
Wash several changes of tap water
Stain with Harris Hematoxylin for 3 minutes Proceed as you would for paraffin  sections Get great staining. 
I am sure if you are using a different Hematoxylin you could use it the same way you use it for paraffin

Donna Reynolds HT(ASCP)
Chief Histology Technician, Core Immuno Lab U. T. M.D. Anderson Cancer Center, Houston, TX
713-792-8106
e-mail dreynold <@t> mdanderson.org

Message: 2
Date: Fri, 12 Aug 2011 08:52:11 -0700 (PDT)
From: Jennifer Sipes <jengirl1014 <@t> yahoo.com>
Subject: [Histonet] H&E for Frozen Tissue
To: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<1313164331.88113.YahooMailNeo <@t> web125406.mail.ne1.yahoo.com>
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Does anyone have a good protocol for staining frozen sections with H&E?? Also, any mounting medium suggestions?? This is the first time I've been asked to do this for frozen. 
?
Thanks a bunch everyone!
Jen
?
Jennifer K. Sipes, ALAT
Sr. Laboratory Technician
Johns Hopkins University
Ross 933
720 Rutland Avenue
Baltimore, MD? 21205
phone:???? 410-614-0131
fax:???????? 410-955-9677
cell:???????? 443-631-6361
e-mail:? jsipes1 <@t> jhmi.edu 

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