[Histonet] Re: Ethanol/other fixations for frozen sections (Andrea T. Hooper)
Well it seems as if there are more then one
answer to this question. Here is what I do. I got this suggestion from
Gayle Callis and it works great for some not all markers.
I section my frozen mouse tissues and set them to dry at room temp
in a hood over night so they are bone dry before fixing. I've also done
this after a few hours and it's just as good. The next day I fix in 75%
acetone/ 25% 100% ethanol for 5 minutes at RT and wash 3x in TBS and go
forward with IHC. C45/B220 works well this way, GL7 did not work well I
saw diminished staining compared to acetone alone. I think Gayle used it
on CD4/CD8 antibodies from BD. The morphology was better then acetone
alone. You may have to test your antibody out before doing your real run.
We had seen this with another reagent MORPHOSAVE, it was hit or miss with
We also have done PFA fixed slide right after sectioning for 2 minutes
then washed in dH20 2x then put in TBS for IHC. The morphology was great
for our CXCL13 antibody from R&D.
Hope it helps.
Sr. Research Associate
Abbott Bioresearch Center
100 Research Drive
Worcester, MA 01605-4341
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