Hi Histonetters,

We are working with mouse and rabbit trachea and are experiencing problems
with the histology. We are seeing fragmentation, lifting off and ultimate
destruction of the epithelial and underlying layers. This does not happen
with all samples but is occuring far too often. We are fixing the mouse
tissue in formalin 18-24 hrs at 4C and then transferring to 70% ETOH. The
rabbit tissue is fixed in formalin at 4C for 24 hrs and then processed. We
are using alcohols for dehydration and xylene for clearing. I see where
other folks are using different dehydrants etc for mouse tissue but this is
not an option for us at the moment. We have processed for very short
durations and longer but still see quite a bit of variation. We are doing
routine staining of the tissue as well as some immunohistochemistry.

Do you have suggestions for fixatives and length and temperature of
fixation?
Suggestions for processing protocols, ie length, temp use of vacuum?
Suggestions for ideal paraffin?

Thanks,
Helen Carney
VAMC/UofU
hcarney@burgoyne.com