Fix for zfish & GFP?

Beth Ann Sipple ayla at astro.ocis.temple.edu
Mon Feb 24 16:47:18 EST 1997


In article
<Pine.SOL.3.91.970220113139.29933A-100000 at teer20.acpub.duke.edu>,
ylin at ACPUB.DUKE.EDU (Yiing Lin) wrote:

> Hello,
> 
> I'm working with GFP (with the S65T mutation) in zebrafish and was 
> wondering if anyone has found a suitable fix that keeps the GFP signal 
> "in place" and doesn't diminish the signal?
> 
> I've tried 4% paraformaldehyde (in PBS for ~2 hrs.) as a fix, but found
> that much of my GFP signal was quenched; what signal was left seemed like 
> it had dispersed a bit into tissue that surrounded the original 
> fluorescent spot.
> 
> Thanks!
> 
>                                                 -Yiing Lin
>                                                  ylin at acpub.duke.edu

When I fix, I prefer to use 4% PF in PBS over night in the refrigerator. 
Sort term fixation at room temperature did not do as good a job.  For
zn-12 antibody stains, 10% PF in PBS also worked well.  Good luck!

Beth

-- 
Beth Ann Sipple
Dept. of Biology




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