Hi Julie:
You mentioned that you are buying your fixative from a commercial
supplier but that
"My first thought is that there was difference in formalin... but that is not the
case."
How do you know? You did not make the fixative solution so you really have no
idea. Not to berate Fisher Scientific, I have been a loyal customer since grad
school. Maybe someone had a bad day or was interrupted during the mixing.
Mixing buffered formalin in you lab is so easy and you retain control of the
most important step in histology.
Geoff
Randolph-Habecker, Julie wrote:
Folks,
I need some input on a problem we're seeing with some mouse brain
samples. The samples are from new born mice P0 to P14 which have been
fixed in 10% NBF (Fisher brand and well within expiration date) for 72
hours. They are then transferred to 70% etoh and processed on an 8 hour
process. After the tissue is embedded, we are taking sagital sections,
drying them overnight, and then baking overnight at 60C. They are then
stained with H&E. This has been working great but now all of a sudden we
have very light H&E staining, nuclear bubbling, and extensive tissue
cracking. I also have noticed some formalin pigment.
My first thought is that there was difference in formalin or time in
formalin but that is not the case. I also wondered if the tissue might
be exposed to excessive heat. We checked all of the temperatures in our
processor, embedding center, and water baths - all were within
tolerance.
Any ideas what might cause all three artifacts? Could it be from
inadequate paraffin infiltration?
Any help would be greatly appreciated!
Thanks,
Julie
Julie Randolph-Habecker, Ph.D.
Staff Scientist - Director
Experimental Histopathology Shared Resource
Fred Hutchinson Cancer Research Center
1100 Fairview Ave, N. DE-360 (Please note new location)
Seattle WA 98109-1024
206-667-6119
jhabe...@fhcrc.org
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcaul...@umdnj.edu
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