Re: H&E on 50um brains

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From:"Barry Rittman" <>
Date:Thu, 19 Aug 1999 06:41:22 -0700
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		the main problem with thick sections is that uneven staining results from
using the routine staining procedures due to the thickness of the sections.
For thick sections we have always used dilute solutions of both the
hematoxylin and the eosin. Hematoxylin (we use Ehrlich's) can be diluted to
1-5% with glass distilled water, filtered and used as is. Staining times
may be hours to overnight but as the stain is so dilute the staining is
progressive and uniform. The solution should be changed a few times and
must always be made fresh and filtered. Differentiation in 0.25% acid
alcohol for shortest time possible to clear up the small amount of
background that results.
Eosin... dilute to 0.25% and add a few drops of 2% acetic acid per 100 ml. 
Staining usually takes several minutes  and again differentiation should be
Having said that I must admit that while we have used this on thick
sections of several tissues we have not done this specifically on brain,
however see no reason why this would not work.


At 11:08 AM 08/18/1999 -0400, you wrote:
>Hi histonetters,
>Does anyone out there have a good procedure for doing an H&E on a 50um
>frozen section of a brain?  Right now, after fixation, we're starting in
>water doing our normal routine.  Everything is quite muddy, however, but
>because of the thickness of the section, I'm not sure it can be any clearer.
>I'd just like to hear what other people are using for their staining times.
>Thanks for any help!
>Emily Yandl

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