[Histonet] slides for frozens

[Sherwood, Margaret]

That may be true, but I don't recall having this problem with our previous
protocol (which was longer and included a fix in alcoholic formalin).  I think
we will try the "old" method and see what happens. 
 

Peggy Sherwood 
Lab Associate, Photopathology 
Wellman Center for Photomedicine (EDR 214) 
Massachusetts General Hospital 
50 Blossom Street 
Boston, MA 02114-2696 
617-724-4839 (voice mail) 
617-726-6983 (lab) 
617-726-1206 (fax) 
msherwood <@t> partners.org 

 

________________________________

From: Rene J Buesa [mailto:rjbuesa <@t> yahoo.com] 
Sent: Thursday, February 02, 2012 2:57 PM
To: histonet; Sherwood, Margaret
Subject: Re: [Histonet] slides for frozens


There is a known saying that goes: "You get what you pay for".
René J.

--- On Thu, 2/2/12, Sherwood, Margaret <MSHERWOOD <@t> PARTNERS.ORG> wrote:



	From: Sherwood, Margaret <MSHERWOOD <@t> PARTNERS.ORG>
	Subject: Re: [Histonet] slides for frozens
	To: "histonet" <histonet <@t> lists.utsouthwestern.edu>
	Date: Thursday, February 2, 2012, 2:46 PM
	
	
	To all:
	
	We have been using the same slides for paraffins and frozens.  We have
no
	trouble with paraffin-embedded tissue adhering to the slides through the
whole
	staining procedure.  But our frozens (even 5um sections)have been
falling off
	during manual staining.  The slides are (+) charged.  We used to use
slides from
	Fisher, but they are so expensive, we changed to an independent vendor
(half the
	price).
	
	Our procedure for frozen H&Es has recently changed:
	1. Fix in 95% ETOH       1 min.
	2. Hematoxylin           5 min.
	3. Acetic acid H2O     2 quick dips
	4. Wash-running H2O      10 seconds (10 dips)
	5. Bluing reagent           10 dips
	6. Wash-running H2O      10 seconds (or 10 dips)
	7. Eosin                 5 seconds
	8. Dehydrate-95% ETOH    10 dips each
	9. Dehydrate-100%ETOH    10 dips each
	10.CitriSolv             10 dips each
	11.Coverslip
	
	The tissue seems to fall off in the bluing reagent (we order from
Mossberg).
	
	Can anyone help us?  Suggest anything--what slides you are using?  We
have had
	the same problem with special stains (i.e. Oil Red O for fat on
frozens). 
	
	Thanks!
	Peggy
	
	P.S.  I use the same slides for my 1um plastic embedding which I dip in
water
	rinses and have no problem with the tissue falling off.
	
	
	Peggy Sherwood
	Lab Associate, Photopathology
	Wellman Center for Photomedicine (EDR 214)
	Massachusetts General Hospital
	50 Blossom Street
	Boston, MA 02114-2696
	617-724-4839 (voice mail)
	617-726-6983 (lab)
	617-726-1206 (fax)
	msherwood <@t> partners.org
<http://us.mc1621.mail.yahoo.com/mc/compose?to=msherwood@partners.org> 
	
	
	
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