what type of tubes are you catching your cells in? Polystyrene by chance? I've seen the catch tubes get a charge built up on them and actually deflect the droplet away. Now this was on an Elite, with much lower droplet velocity. I figure if something isn't making sense, you have to check everything. Are you getting a droplet forming on the rim of the catch tube? That's the telltale sign. On Tue, 29 Apr 2008 14:32:29 +0200, Simona Ronzoni wrote: >Dear Flowers >I have a problem with FacsAria >When I sort stem cells from KASUMI cells linee (Acute >MyeloId Leukaemia), I lost the cells. >My guess is that the cells are being lost due to stickiness or cell >death. >Looking at FSC vs. SSC and gating on the viable cells post sort only >10% of the events fall in what I would consider debris or dead cells. The >cells are spin down and then counted. There are essentially no dead cells >visible >in these counts. >Last time I sorted 3.200.000 cells and I have count only 11.000!! It's >impossible!! I don't have the same problem with other cells type and I use >always the same parameters ( PSI, Drop delay and Frequency) >Can you help me? >Any insights will be greatly appreciated. >Ciao >Simona >-- >Simona Ronzoni >Imaging Centre >Consortium for Genomic Technologies >c/o IFOM-IEO Campus for Oncogenomics >via Adamello 16 >20139 Milan >Italy >Phone: ++39-0294375087/0294375038 >email: simona.ronzoni@ifom-ieo-campus.it >http://www.ifom-ieo-campus.it/RESEARCH/Imaging/ >http://www.ifom-ieo-campus.itReceived on Mon May 5 15:38:00 2008
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