Hello Siow Fong Lee, Cells travel through the cytometer in a narrow core stream that is made up of whatever the fluid is in the sample suspension. This narrow stream is contained within a wider stream -- that is the fluid in the sheath reservoir. This is usually phosphate-buffered saline. So the amount of the sample fluid/lysate that comes through in the sort is dependent on how wide the narrow core sample stream is. This width is dependent on the configuration of your cytometer and also on how hard you push the cells to get them to flow. If you use high pressure, then the central sample core stream is wider and more of the lysate will get sorted along with your cells. Mostly the narrow sample core is somewhere around 5-10 microns in diameter. Usually the sheath stream is about 70-100 microns in diameter. If this is the case, then the total volume that gets sorted with your cells will be about 1% sample lysate and the rest sheath reservoir 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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