fixation penetration

William P Irwin wpirwin at titan.oit.umass.edu
Thu Oct 26 17:33:12 EST 1995


Nancy Bray (nbray41 at MAINE.MAINE.EDU) wrote:
: I am looking for ideas/methods to increase the permability of Dros. cuticle
: prior to or during fixation for light microscopy and electron microscopy.
: Any suggestions welcome.... I want to fix these critters intact.
: Thanks
: Nancy Bray
: nbray41 at maine.maine.edu

I needed to have cuticle penetration for x-gal staining and found that 
50% glutaraldehyde on ice for 3 minutes worked wonderfully (followed by 
PBS washes, etc.).  I normally use 0.2% glut. in PBS for fixing nerve 
tissue that has been dissected and thought that using 50% was kind of 
whacked, but I found a reference to the method in a paper, I think by A. 
Ghysen (I'll look for it and get back to you) and tried it since nothing 
else was working.

Hope this helps,

-Bill




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