How to get through cuticle?
Tritech_Research at lamg.com
Fri Jul 26 13:41:54 EST 1996
Susan Jane Hogarth writes:
I'm looking for a good way to get through the worm cuticle, leaving
(opimally) nice, intact tissue. Any ideas? Chemical digestion?
sonication? Grinding in N2? ???
Thanks for the help.
Since a few people have posted asking about this, I have a couple of
observations to share, though I am by no means an expert about this.
We got through the cuticle well enough to get good B-gal staining by
simply putting worms on a microscope slide (in a drop of water),
letting the water evaporate (can be accelerated w/ a heat lamp, etc.),
and repeatedly micropipetting drops of acetone on them. (This was
pretty much a suggestion of Andy Fire's. ) If you monitor this under
the dissecting scope, you will see the worms get get all desiccated,
until the become what I would call birefringent (but I'm not sure
if I am using that word correctly). At that point, they are very
collapsed in on themselves and presumably the cuticle, etc. gets
busted up. When you rehydrate with an aqueous solution, the worms
return to an amazingly normal appearance, but are quite permeable.
Another thing that permeates the cuticle efficiently is electroporation,
at least when using our cool hand-held electroporator, the Cloning Gun.
TRITECH_RESEARCH at LAMG.COM
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