Hanging drop method for C. elegans embryos
In this protocol, we show how to prepare C. elegans embryos on a standard coverslip to perform experiments with CherryTemp using the hanging drop method. It shows step by step how to dissect and mount the sample for the experiment.
Step 1 : Pick young adults into pre-cooled M9 medium (16°C).
Step 2: Dissect them by cutting at both ends of the uterus using a scalpel and #15 blade to release the embryos.
Step 3: Choose the embryos of the desired stage and transfer them to a cover slip using a mouth pipette.
Step 4: Use a syringe 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.
Step 5: Use a mouth pipette to transfer the embryos, cluster embryos with an eyelash tool.
Step 6: Carefully cover with our temperature control chip. Gently clamp the system using our insert.
Step 7: Place the mounted slide on either an upright or an inverted configuration by simply rotating 180°.
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