Use our dedicated spacers to mount C. elegans temperature-sensitive embryos
Introduction
This protocol shows how to mount C. elegans embryos on glass coverslips with the CherryTemp system. Spacers of different thicknesses create chambers where embryos are placed. Spacers can replace standard methods like hanging drop method on glass coverslip.
Step-by-step guidelines:
Step 1: Pick young adults into pre-cooled M9 medium (16°C).
Step 2: Dissect them by cutting at both end 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: Top the coverslip with our dedicated spacer.
Step 5: Place the live embryos on the spacer hole, in a volume of up to 15ul.
Important: medium must not touch the sides of the spacer.
Step 6: Top with the CherryTemp chip (engravements facing up, coverslip and liquid microfluidic chamber facing the sample).
Step 7: Top with the CherryTemp chip (engravements facing up, coverslip and liquid microfluidic chamber facing the sample).
References
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
