Posted by
Glen MacDonald-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/C-elegans-imaging-tp6826291p6832322.html
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Hi Claire,
Definitely use glass bottom plates, but they are expensive. We use them for screening zebrafish with 10X/.4 or .5 (3 mm wd) and 20X/.75 lenses (650 um wd). We re-used them many times after bleaching. Maybe a hot water treatment to remove the agarose. Can defined amounts agarose be added by a multi-well pipettor to better control thickness? If you are restricted to a ~1 mm agarose thickness, the maybe a high mag, low NA lens or a place a doubler in front of the camera to achieve Nyquist.
Regards,
Glen
Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923 USA
(206) 616-4156
[hidden email]
On Sep 23, 2011, at 6:37 PM, Claire Brown, Dr. 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