egg permeabilization

Michael Finney mikef at mjr.com
Fri Dec 22 18:05:34 EST 1995


>I am working to try to fluorescently label the egg cell nuclei of
>Heterodera glycines eggs so that I may evaluate the developmental stage
>of egg populations using flow cytometry.
>
>Chris Marett
>cmarett at iastate.edu

Chris,

You didn't say whether it is OK to kill the eggs in the process. I'll
assume it is since you're doing flow cytometry afterward.

I know of two treatments sufficient to permeabilize C. elegans eggs with
high efficiency, though not everything in these protocols may be actually
necessary. They are:

1 hypochlorite treatment (has the advantage of getting rid of all tissues
except eggs) followed by formaldehyde fixation.

2 formaldehyde fixation accompanied by a freeze-thaw cycle.

For more information, check out this site on the web:

gopher://weeds.mgh.harvard.edu:70/00/caenorhabditis/mgh_protocol

There are two protocols there that might be of interest, one on
hypochlorite treatment and one on fixation and permeabilization.

Mike

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