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Hanging drop method for C. elegans temperature-sensitive embryos

Hanging drop method for controlling C. elegans temperature-sensitive embryos

1/ Early-stage embryo dissection

Pick young adults into pre-cooled M9 medium (16°C).

Dissect them by cutting at both end of the uterus using a scalpel and #15 blade to release the embryos.

Choose the embryos of the desired stage and transfer them to a cover slip using a mouth pipette.

Early-stage C. elegans embryos dissection for live-imaging analysis

Early-stage C. elegans embryos dissection for live-imaging analysis

2/ Mounting using a Vaseline ring

Use a syringae to make a ring of Vaseline on a coverslip. Make sure to center the ring so that the embryos will be correctly placed under the thermalisation pattern of our chip.

Use a mouth pipette to transfer the embryos, cluster embryos with an eyelash tool.

Carefully cover with our temperature control chip.

Gently clamp the system using our insert.

Schematics of the hanging drop method

 Methods adapted from J. Dumont and J.C. Canman:

T. Davies, S. Sundaramoorthy, S.N. Jordan,M. Shirasu-Hiza, J. Dumont and J.C. Canman, 2016, “Using fast-acting temperature-sensitive mutants to study cell division in Caenorhabditis elegans“, Methods in Cell Biology, Volume 137, ISSN 0091-679X, http://dx.doi.org/10.1016/bs.mcb.2016.05.004 

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