1.)Attempting to use all the laser lines simultaneously has not been easy. I try to resolve 8 peak rainbow beads (Spherotech)for the UV using a 450/65 BP, the peaks are all scrunched together. I attempted to use a 488 blocker with the 450/65 BP, because I thought the 488 was causing the problem, but ended up loosing all my signal. I'm wondering if a 488 shortpass would work in front of the UV pathway to keep out 488 from bleeding through. Does anyone have any ideas? 2.)I'm also interested in finding a bead that I can run after setting the PMT voltages and parameter gains with unstained cells. I'd like to have a fluorescent particle control that I run at the beginning and end of each set of samples, utilizing all the settings I run the samples under, to ensure my customers the machine is not causing any anomalies that may arise from time to time in the experiments. I would like to have a negative bead and a positive bead that is excited by a UV line, 488 line, and 633 line. Can anyone offer me any advice? 3.) Lastly, I'm looking for an alignment bead for when I set-up to run APC-Cy7. Does anyone know of a bead that emits at 767 nm and excited by a 633nm laser line? Thanks, Howard Howard Robinson Flow Cytometry Core Facility Joslin Diabetes Center Room 491 One Joslin Place Boston, MA 02215 Howard.Robinson@Joslin.Harvard.Edu (617)-732-2400 (Ext. 4516)
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