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Fluorescence Microscopy Woes

Glenn White gwhite at ashland.edu
Fri Oct 23 10:56:29 EST 1998


Howdy:
We're having some problems following the Finney/Ruvkun protocol in David's
article in the Worm protocol book.

Our problem is that we not getting good permeabilization of adults and
they are not taking up much phalloidin label.  Younger animals show a
higher percentage of staining but it's not like 90+% percent.

We're freezing and thawing the worms at least 3 times during the 1 hour
fixation period.  All of our reagents are fresh from the company.  The
peroxide was just purchased two weeks ago and has only been opened 2 times
so far.

I've gotten this protocol to work beautifully before (but before I came
here), so I'm a bit perplexed.

Any suggestions or comments would be most appreciated.

Cheers,

Glenn

Glenn Edward White, Ph.D.
University Honors Program Director
Associate Professor of Biology
Ashland University
Ashland, Ohio 44805
419-289-5259
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