[Histonet] Re: Ethanol/other fixations for frozen sections (Andrea T. Hooper)

[Jamie E Erickson]

Hi Andrea,
                          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 
some antibodies. 

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.

Jamie

_______________________________
Jamie Erickson
Sr. Research Associate 
Department: DSMP
Abbott Bioresearch Center
100 Research Drive
Worcester, MA 01605-4341
508-688-3134
FAX: 508-793-4895
e-mail: jamie.erickson <@t> abbott.com