hello! I have taken upon myself to run some calibration and performance tests on our institution's old and venerable FACScan. Luckily the person actually in charge of the FACScan and the Vantage (SE) is very nice and wants to cooperate. Any ideas, tips, tricks, suggestion etc are VERY welcome. 1)We'll run single-intensity beads to determine CV, which I understand also gives a measure of the alignment of the laser. How much CV is acceptable? 2)Using 6 peak fluorospheres from Dako, I'll perform the experiment detailed on HS PFC 4th ed pag 201-202 to test log am linearity: I'll put the highest peak at or close to the top channel, acquire, and then lower the PMT voltage to put the highest peak 5-10 channels lower, acquire, etc and so forth. This will be repeated for FL1, FL2 and FL3. I am assuming that the lower 3rd of the lowest decade and the upper 3rd of the highest decade will be bad, but I do want 3 1/2 decades of dynamic range! How much deviation from the linear response is acceptable? should I perform it using 256 or 1024 channels? 3)Are there any other (simple) tests to measure the FACScan's performance? which parts need to be tested? 4)PFC states that noise can be easily 3 mV, and once again assuming this FC has been neglected a bit, how can I determine the noise at the lower scale? 5)PFC says that below 300 volts the PMT starts to behave non-linearly. Given that these PMT are old (they seem to have been replaced since it was bought, but not sure when), I was thinking maybe they are not behaving well even at higher voltages. How can we measure their performance? Is this test needed or not? 6)Assuming the alignment test, the log amp test, or other tests result unacceptable, is there something to be done by us, or the only way is to call the BD technician? Part of my effort here is that in case the FCytometer is not performing well, show them, so that they will be compelled to place a maintenance call, since it has been long since it was done last time. You know, "if it's working, why does it need repairs?". If the tests are ok, even better! They do have Current protocols in FC, but I haven;t had the time to sit and read the relevant chapters, although I'll do it before getting to work. Thanks in advance, Uriel.Received on Mon Sep 27 15:18:00 2004
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