An investigator here wants to sort C. elegans embryos expressing GFP. The embryos are anywhere between the 28 cell to 600 cell stage with most closer to the smaller size. Each embryo is roughly 50x30 microns, ellipsoid in shape with a fairly rigid shell encasing the embryo. We are told that the embryos are relatively resistant to physical trauma. We have run these on a Vantage with a 100 micron tip (no Macro Sort) and can detect the GFP expressing embryos. Our problem arises with the sorting; if we sort 40,000 GFP positive embryos, one only finds about 100 in the collection tube. The good news is that those 100 are healthy in that they grow normally following the sorting process. The question is, what might cause this abysmal sorting efficiency? When we sort "normal" cells like murine stem cells, lymphocytes, etc. we get very good recovery and purity. If there is anyone out there with experience sorting something like this I would greatly appreciate your suggestions. Thanks Wayne F. Green, Ph.D. HCI Flow Cytometry Core Lab wayne.green@hci.utah.edu
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