Michael G. Ormerod Michael_Ormerod at
Mon Aug 24 17:25:16 EST 1998

I would look at the cells under a fluorescent microscope after they have
been trypsinised off the substrate. You will then be looking at them in the
same condition as the flow cytometer.

When you run cells through the flow cytometer, it collects fluorescence
from the whole cell. When you examine the cells by eye, your perception of
brightness tends to be governed by the fluorescence of the brightest areas.
If an equal amount of EGFP is distributed differently in two types of cell,
you may observe an apparent discrepancy between the eye and the flow
cytometer. For example, if the protein in one cell is concentrate in a
particular organelle, it may look quite bright. The same amount of protein
distributed evenly through the cytoplasm could appear quite dim, you may
not even be able to see it.

Michael Ormerod
34 Wray Park Road
Reigate RH2 ODE
Telephone: voice & FAX: (0)1737 241726
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