Re: C. elegans imaging

Posted by Cliff Luke on
URL: http://confocal-microscopy-list.275.s1.nabble.com/C-elegans-imaging-tp6826291p6829064.html

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Hi Claire,

Currently we use optical bottom 96 or 384 well plates for our HCS imaging of C. elegans
but we have the luxury of the worm biosort to dispense the worms into the wells. Another
way we have been doing screens is to wash the worms off the plate with PBS + o.o1%
triton x-100 (so the worms don't stick to the side of pipet tips etc.) let them settle on ice
then manually dispense into wells of the optical bottom plates. Current anesthesia we use
is 0.05M NaAz or 1mM levamisole.

If you want to keep the on the 24 well plates then you could try using more NGM filling
the well up and the using optical sealing film and turn the plate over. Not great for bright
field but an option, however they will move like crazy. The woes of C. elegans imaging.

Let me know if I can be anymore help

Cheers

Cliff

Cliff Luke, Ph.D.
Assistant Professor, 
University of Pittsburgh,
Department of Pediatrics,
Children’s Hospital of Pittsburgh
One Children’s Hospital Drive
4401 Penn Avenue
Rangos, Rm 7131
Pittsburgh, PA  15224
USA

Tel: [phone]
Fax: [phone]
email: [hidden email]

On Sat, 24 Sep 2011 01:37:24 +0000, Claire Brown, Dr. <[hidden email]>
wrote:

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>
>
>We have a lab that wants to screen a C. Elegans genetic library with about 18,000
genes. They want to take images of the worms on 24 well plates and measure the
distance between 2 of 3 EGFP spots on the worms.
>
>We would like to do the experiments on an inverted HCS automated microscope. The
problem is they have the worms growing on a ~1 mm thick nutrient-rich agar layer.
>
>We need to image the worms through the agar so we need to find a different way to
prepare the samples. Has anyone ever tried something like this? If so do you have a
solution for this? The agar is poured by hand so getting it much thinner than 1 mm would
be difficult.

>
>
>Sincerely,
>
>Claire
>
>--------------------------------------------------------------------------------
>Claire M. Brown, PhD
>McGill University
>Life Sciences Complex Imaging Facility Director