Re: Hematoxylin & Eosin Staining on Frozen Brain

From:"J. A. Kiernan" wrote:
> I am trying to find an H&E protocol for staining frozen brain sections cut at
> 7 microns. I have tried to stain the samples using the same protocol I use
> for other tissue types but I am not getting good results. If anyone has any
> advice, I would really appreciate it.

If the frozen sections are of unfixed tissue you should 
fix them before staining. Alcohol or an alcohol-acetic
mixture for a few minutes should be OK. 

If you really want your frozen sections to look like 
paraffin sections of formaldehyde fixed tissue, immerse 
in formaldehyde for 12-48 hours, wash, dehydrate in 
100% alcohol, and rehydrate before staining. 

After thorough formaldehyde fixation cell nuclei 
look less interesting than after a coagulant fixative, 
which generates characteristic chromatin patterns in 
different cell types. (The boring formaldehyde nuclei
probably more closely resemble the pre-death cellular
architecture!) Eosin staining is weaker after adequate
formaldehyde fixation than after a simple coagulant
fixative, but it's easy to adjust this counterstain.

With regard to brain, it's important to know that
_H&E is a lousy stain for the CNS_ even after ideal 
fixation etc. There are lots of better methods, and 
many of them are easier to do than H&E. 
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1

<< Previous Message | Next Message >>