RE: [Histonet] Cell Culture Staining

From:"Davis, Gareth"
[Histonet] Cell Culture Staining
When I H&E my frozen sections I don't use Xylene.  I merely skip the Xylene and alcohol steps, go straight to dH2O and stain.  When I get to Eosin, I just rinse with dH2O and mount coverslips on (if possible) with ageous mounting media.  This is probably no help if you have no way to coverslip them.  But, this is my suggestion.
Gareth
 
Ms. Gareth B. Davis
Research Assistant II
Neuro-magnetics Division of
the Department of Neurology
Vanderbilt University Medical Center
615-936-3318
 
 
-----Original Message-----
From: Johnson, Kevin [mailto:KJohnson@med.miami.edu]
Sent: Fri 10/3/2003 4:29 PM
To: 'histonet@lists.utsouthwestern.edu'
Cc:
Subject: [Histonet] Cell Culture Staining

Greetings, all.

A researcher just brought me (at Miller Time on a Friday!) two culture
flasks of terminally differentiated Sertoli cells and requested H&E
staining.  Well. 

Conventional H&E obviously is out, since polystyrene is not compatible with
xylene.  Further restrictions: the cells cannot be scraped off, they cannot
be replated on glass coverslips, and for reasons of her own, they cannot go
back into the incubator pending an answer.  They now have been fixed in situ
with 10% neutral buffered formalin.

Does anyone have a protocol for staining these puppies?  If not H&E per se,
then is there another staining method that might satisfy her?

Thanks in advance,

Kevin Johnson
University of Miami
Diabetes Research Institute


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