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Fluorescence Microscopy Woes

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

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.



Glenn Edward White, Ph.D.
University Honors Program Director
Associate Professor of Biology
Ashland University
Ashland, Ohio 44805
"I love being alive and I will be the best man I possibly can.  I will
take love wherever I find it and offer it to everybody who will take it.
Seek knowledge from those wiser, and teach those who wish to learn from me."
					Duane Allman

"Emancipate yourself from mental slavery, none but ourselves can free our
minds."					Bob Marley

"Do not seek to follow in the footsteps of the wise. Seek what they
sought."				Basho

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