Hello Tony, A few ideas to get you started. It sounds as if there may be problems with the threshold set at the wrong level. It may be that you see nothing going through the instrument because the threshold is set too high. You could try setting it lower, and then see if you start to get red blips on the instrument panel. That is always my first guess when the fluid level is decreasing but there appear to be no cells. Water always gives a much higher background flow rate than buffer (amazingly high) --- because the refractive index of water is different from that of the sheath fluid and, therefore, a lot of stray light twinkles off the water buffer interface. The flow rate should be lower if you go to clean, filtered sheath fluid. Alice Alice L. Givan Englert Cell Analysis Laboratory of the Norris Cotton Cancer Center Dartmouth Medical School Lebanon, New Hampshire NH 03756 tel 603-650-7661 fax 603-650-6130 givan@dartmouth.edu
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