Dear experts in flow sorting: I am trying to sort HUH7 cells GFP (brigth and not so bright populations) Just two gates in a single color FL1 vs SSC (very simple sorting should be). Those are big cells so I tried with the 70um nozzle but with medium pressure. I haven't had problems with settings, or with accudrop. However, I couldn't have pure populations. When setting the streams I can see the two perfect dots on the camera on the screen and accudrop run perfectly, but when I am sorting I don't see but a smear of white dots to the right and to the left of the main stream (is that because of the big cells?). I am planning to try the 100um nozzle, but can I run the accudrop with this nozzle eventhough they are very small? If somebody can give me some tips, I will be very grateful. And another question, if I prepare the machine for asceptic sorting, can I run the accudrop before or I need to prepare a sterile solution of these beads or run them after asceptic cleaning and then run again bleach and water for a few minutes to keep the "sterility". (We do not have the FACSAria in a BSL-3 lab yet) Thanks a Lot VidaReceived on Wed May 7 13:38:00 2008
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