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You may want to look up the following (old) reference. We were labeling
HLA-DR and MAC-1 on human monocytes-on-beads. We found that with a
great deal of fooling around we could fairly reliably get a more-or-less 1:1 ratio
of beads to cells, but as I recall, we never got as much labeling of the
cells-on-beads as we cold get on cells-in-suspension. I always thought that
perhaps most of the adhesion molecules on adherent cells are localized on
the adherent side and thus may not be accessible to antibodies in solution. I
don't know how you could get your label between the bead and cell - we did
EM and found not much space there. Also, it becomes somewhat of a
philosophical issue as to whether the cell thinks it is adherent to the bead or it
thinks it has eaten it! We weren't sure if were were studying adherence or
phagocytosis. I don't recall what sized beads we ended up using. If they are
too big you get many cells per bead and then of course they won't "flow". It
they are too small you get many beads per cell and you are definitely studying
phagocytosis.
Bloch, D.B., B.R. Smith, and K.A. Ault. 1983. Cells-on-microspheres: A new
technique for flow cytometric analysis of adherent cells. Cytometry 3:449-452.
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Kenneth A. Ault M.D.
Maine Medical Center Research Institute
125 John Roberts Road
South Portland ME 04106
Voice (207) 761-9090
Fax (207) 761-2130
E-Mail aultk.mmcri@office.mmc.org
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