Dear Geoff, When I was setting up my CloneCyt I was playing with fluorescent beads a lot. If you can't take plate to the fluorescence microscope, put a slide over it and then sort. Even when drop dries out you should see beads with no problems. From my experience, Vantage is very reliable in sorting 1 particle. Take 3 drops envelope (effectively it's 4 for the CloneCyt, adds 1/2 drop both sides, just in case), go slow rate (if it's too fast you could not have single target in 4 drops at all) and you should not miss out your cell, although you are going to waste relatively lots of them while doing this. I presume you did a few drop delay profiles before, that you can trust to. Hope this helps, Sasha. •••••••••••• Dr Sasha Sreckovic Dept Path & Micro University of Bristol University Walk Bristol, BS8 1TD, UK Sasa.Sreckovic@bristol.ac.uk 0117-928-8606 •••••••••••• ---------- >From: "geoff.morgan" <geoff.morgan@bbsrc.ac.uk> >To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> >Subject: CloneCyt/Single Cells >Date: Tue, Feb 1, 2000, 12:17 pm > > > Dear Flowers,I have recently started using CloneCyt on a FacsVantage.We > have benn sorting single cells into a 96 well plate which then has > PCR. Cells are usually detected in half the wells.When I sort 5 cells > onto a lid I can usually find 5 cells in most positions(by dissecting > microscope), finding single cells is a lot harder. I intend to try m> again using blue stained cells.What sort of success do other people get > any ideas how I can check/improve it Thanks Geoff Morgan Babraham, > cambridge U.K. > > geoff.morgan@bbsrc.ac.uk > > > > > > > > > >
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