[Histonet] Re: H&E staining of frozen section
Steven E. Slap
siksik03 <@t> comcast.net
Tue Apr 27 14:52:42 CDT 2004
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 55°C 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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