[Histonet] slides for frozens

[Rene J Buesa]

That could be the solution. The problem is that you changed 2 things: the protocol and the slides and now you do not know which is "the culprit". Returning to the old protocols is a first step to trying to solve the present situation and, as you can imagine, if the old protocol does not solve the problem, the slides may be the cause.
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: "Rene J Buesa" <rjbuesa <@t> yahoo.com>, "histonet" <histonet <@t> lists.utsouthwestern.edu>
Date: Thursday, February 2, 2012, 3:03 PM


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> 
    
    
    
    The information in this e-mail is intended only for the person to whom
it is
    addressed. If you believe this e-mail was sent to you in error and the
e-mail
    contains patient information, please contact the Partners Compliance
HelpLine at
    http://www.partners.org/complianceline . If the e-mail was sent to you
in error
    but does not contain patient information, please contact the sender and
properly
    dispose of the e-mail.
    
    
    _______________________________________________
    Histonet mailing list
    Histonet <@t> lists.utsouthwestern.edu
<http://us.mc1621.mail.yahoo.com/mc/compose?to=Histonet@lists.utsouthwestern.edu
> 
    http://lists.utsouthwestern.edu/mailman/listinfo/histonet