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Hi Claire,

It depends on the working distance of the objective you are using. Some
objectives can focus on a specimen a few millimetres away from the lens,
so 1 mm agar on top of a plastic plate wouldn't be a problem. But of
course you'll have reduced NA for longer WD.

It also depends how they prepare samples. Do they need to image moving
worms? If so, they will need some kind of worm-tracking system which
could complicate things more. If the worms are somehow paralyzed for
imaging, then I don't see any reason why agar has to be used. You can
keep them in liquid for a while.

In my case, I place worms in between a cover slip and thin agarose layer
to minimize the working distance and disturbance of worm's locomotion.
With this setup, relatively high NA objective (0.75) can be used for
brighter images of moving worms.

One other note; I use agarose instead of agar because agar has too much
auto-fluorescence.

Best,

Ippei


(11/09/23 21:37),
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
> 
> 
> 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
> 


-- 
Ippei Kotera, PhD

Centre for Research in Neurodegenerative Diseases
Tanz Neuroscience Building, Room 221
University of Toronto
6 Queen's Park Crescent West
Toronto, Ontario M5S 3H2

Phone: (416)978-2503
Fax: (416)978-1878