Artifact: poor fixation or inadequate slide drying
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From: | Adrienne Vair <a_vair@hotmail.com> |
To: | histonet@pathology.swmed.edu |
Reply-To: | |
Content-Type: | text/plain; format=flowed |
Our pathologist's are concerned with the poor nuclear morphology they have
been routinely seeing for many years; foamy appearance, muddy nuclear
staining (H&E). I suggested that we look at extending the time in 10%
formalin as up until then, surgical specimens were received throughout the
day from the O.R. in 10% Neutral Buffered Formalin (NBF) and most were being
processed the same day with the exception of bowel, uterus, and breast
tissue which were being opened and fixed 24 hours. We extented the fixation
time to a minimum of 24 hours for all surgical specimens including
colposcopy, endoscopy and fine needle biopsies but to no avail (and the
increase in turn around time concerned the physicians). The morphology
remained poor. I looked at temperatures of all reagents during processing
and embedding, and checked the pH of the 10% formalin being made up and used
routinely. I stopped using the microwave oven to dry slides as I could not
find any literature to confirm that microwaving a rack of 20 slides for 7
minutes at full power (with small containers of water in the oven chamber)
did not affect the tertiary structure of the proteins in the tissues. Nor
could I find a procedure in the lab or references as to where the idea
originated. They are now being dried at 60 degrees celcius for one hour in
an oven prior to staining. The pathologist's insisted that the problem was
not with fixation but with staining, I made improvements there but as I've
stated before I could not improve on the nuclear morphology. My question is,
is this a fixation problem? Is this a slide drying problem as I have
recently read on this site? I have read the article on alcohol/xylene
tissue processing (Journal of Histotechnology March 2000 issue page 45, and
the ideas seemed feasible but because it is unconventional, I would
appreciate some other ideas and comments.
Thanks for the input.
Adrienne Vair MLT BMLSc
Histology Supervisor
Medicine Hat Regional Hospital
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