From: Mario Roederer (Roederer@Beadle.Stanford.edu)
Date: Fri May 15 1998 - 11:30:42 EST
Cathy: Back when I was a grad student with Bob Murphy, we used to see 0.3 uM beads readily on a FACS 440. We had to use log SS as the trigger parameter, and use specially processed sheath fluid (we would highly filter it and then spin it in an ultracentrifuge for an hour or overnight to pellet debris). This reduced "background" triggering to about 50 events per second or so; the 0.3 uM beads could be seen as a distinct population above that. However, I seriously doubt that 80 nm vesicles will appear by scatter. However, if you could make them fluorescent, you would have an excellent shot at success. For example, you could load them with nile red (membrane dye), and trigger off of red fluorescence. They should likely achieve enough fluorescence to be easily detected. However, scatter signals will be meaningless (in fact, probably absent) even when triggering on the fluorescence. mr
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