[Histonet] Problems with mouse brains

[Geoff McAuliffe]

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
> jhabecke <@t> fhcrc.org
>
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>
>   


-- 
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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 
mcauliff <@t> umdnj.edu
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