[Histonet] Problems with mouse brains

[Randolph-Habecker, Julie]

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
jhabecke <@t> fhcrc.org