[Celegans] Re: Stop moving, worms!

Sam Smith via celegans%40net.bio.net (by sasmon from gmail.com)
Tue Mar 15 18:35:28 EST 2011


On Feb 11, 2:50 pm, Sam Smith <sas... from gmail.com> wrote:
> Hi again.
>
> Our lab is trying to find a way to take confocal images of live, adult
> worms without them twitching.  So levamisole is out, and sodium azide
> is out as we are specifically looking at ETC function which is
> inhibited by sodium azide.
>
> Any awesome ideas out there?

I wanted to give an update to anyone interested.

I got a few suggestions:
-muscimol
-microfluidic channels
-microbeads

Because of the ease of use, inexpensiveness, and the requirements for
confocal imaging (our intended use), I went with the microbead
suggestion.  The protocol can be found here:
http://www.wormbook.org/wbg/articles/volume-18-number-1/agarose-immobilization-of-c-elegans/

A couple notes:  While this has not worked to completely disable
movement of the head region, the worms are not engaging in pharyngeal
pumping-- and the head movements are intermittent which should allow
for sporadic imaging good enough for our needs.  The bodies of adult
worms are completely still. If you are looking at the Lstages, this
will probably not work; their smaller bodies do not allow for an
adequate amount of friction to keep them immobile. I'm also using up
to 2-3uL of the microbead solution. Our coverslips are the longer
style since the slide will need to be upside down to image. It looks
like water loss will be limited. We still have yet to see if there
will be any interference between the beads and imaging, but we have
our fingers crossed.  If you'd like followup after we use them on the
confocal scope, feel free to shoot me an email.

Thanks to everyone who offered suggestions!




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