Re: [Histonet] Re: H&E staining of frozen section

From:"Steven E. Slap"

Hi HistoNetters

I have suggested this many times in the past, but I'll say it once again...

Nuclear details will be much better preserved if 
the cryostat (frozen) sections are first fixed in 
a microwave (see Chapter 14 of Kok & Boon, 
"Combining microwave and freeze techniques"). 
The easiest way to do this is quickly put the 
slide in a horizontal position on a polystyrene 
platform in the microwave and cover the section 
with a few drops of Kryofix or equivalent 
PEG/ethyl alcohol fixative.  Microwave at 450W 
for 20 seconds.  Alternatively, you can place the 
slide vertically in a Coplin jar filled with 
fixative and microwave at 55C for 20 seconds. 
The slide should still be wet, in any case, after 
microwaving.

Other methods have used one part 40% formaldehyde 
and 3 parts 96% alcohol, or Wolman's fluid (5% 
acetic acid in industrial alcohol), but I prefer 
the PEG/ethyl alcohol methods.

After fixing, stain as suggested by Gayle Callis.

best regards,
Steven Slap
Microwave Consultant


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