Hi, I am measuring calcium responses in neutrophils loaded with Fluo3. I run a sample of just loaded cells to measure the basel level and then I run another sample that I add fMLP to as a stimulus. I have the problem that the fMLP stays in the lines of the cytometer and then stimulates my next basel sample. I have tried several water washes(60s), priming and running cleanse cycles with some success over 5-10min. One thing that works, is to do a complete bleach shut-down. All of these methods are impractical due to the number of samples I need to run and the time it will take. I have heard people mention running DMSO to clean out the lines but I don't know what concentration they us or for how long. I am interested in DMSO because it is the solvent used to dissolve fMLP. If anyone is using a cleaning protocol like this will you please give me the details. If anyone has any suggestions, that would be appreciated as well. Thank you, Beth Whalen UBC Pulmonary Research Lab. Vancouver, B.C.
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