Fixation of Cryostat Tissues

From:Vanessa Heim <heimvk@bu.edu>

Hello Histonetters,

I know this is a very basic question but I really need some advice on
basic histology.  Keep in mind I have had absolutely no formal training
in histology and I am teaching myself the techniques of this
science/art.  Due to the overwhelming support of many members of this
list, I have been able to pretty well master the art of cryostat
sectioning.  I no longer have a problem with the tissues falling off the
slides and I can get consistenely good sections when I section without
wasting the whole day with the cryostat.  This brings me to my current
dilemma.  I am having trouble finding the best fixative for the tissue
after I have done the cryostat sectioning.  I am trying to Masons
Trichrome Statining on Penile cross-sections and I am finding that the
tissues are being damaged in the process.  Since I have perfomed this
staining protocol many times in the past with paraffin-embedded tissues
and have gotten wonderful results...i suspect that the fixation or lack
of it is the problem that is creating such horrible looking slides.
Thank you in advance for all the help you can give me with this
situation.

Vanessa Heim
Research Assistant
Boston Univ




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