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Agar pad method for C. elegans temperature-sensitive embryos

Agar pad 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

2/ Mounting on an agarose pad

Add a small drop of 2% agarose on a 170µm thick cover slip and cover it with a perpendiculary placed slide (not a cover slip!) to make the agarose pad.

Once cooled, twist slides relative to each other to expose the pad. It must remain on the cover slip.

3/ Two possibilities next to fit with our CherryTemp control unit

Add embryos in a 5µl drop of M9, gently add the agarose pad on top. Top with our emperature control chip. Gently clamp the system using our insert.


Directly add embryos, top with a cover slip, turn the slides upside down. Place the “slide sandwich” in our insert. Add a drop of immersion oil and top with our temperature control chip. Gently clamp the system using our insert.

Schematics of embryos on an agarose pad

 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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