[Histonet] C. Elegans fixation

[Philip Oshel]

Danielle,

This is a good question for the microscopy list (subscribe at 
http://www.microscopy.com ). There's probably a C. elegans list that 
would be good to consult.
There are also excellent resources on the web. A particularly good 
one is David Hall's lab, the Center for C. elegans anatomy at 
http://www.aecom.yu.edu/wormem/
There are excellent methods here.
Good luck! Intact C. elegans is non-trivial. The cuticle causes much 
fuss and bother, and most people doing EM use high-pressure freezing 
methods, followed by freeze-substitution embedding.

Phil

>Hello Histo-experts,
>
>
>
>Can you please share your favorite worm fixation methods with me??  I've
>had great success in EM prep using 2% PFA 0.2% gluteraldehyde in
>cocodylate buffer.  BUT-I had to "nick" the worms to get infiltration of
>the fixative.  I now have a user who would prefer to avoid the "nicking"
>technique for his application, but so far, we're experiencing
>insufficient fixation.  Any/all advice is very welcome!!!
>
>
>
>Many thanks in advance!
>
>
>
>danielle
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>
>
>Danielle Crippen
>
>Morphology and Imaging Core
-- 
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576