Re: Sorting Issues

From: Alice L. Givan (Alice.L.Givan@dartmouth.edu)
Date: Tue Aug 13 2002 - 13:44:10 EST


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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