Low level fluorescence

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I am performing flow cytometry analysis on human red cells. Due to the low
number of antigen copies per cell, the resulting fluorescense (FITC) is
close to the flowcytometer (FACScan) noise level.

1. Does anyone know how to calculate the real number of cell-bound FITC
molecules (and hence antigens), when working in this low area of
fluorescence intensity?

2. Unmarked human red cells have a lower green (FITC) autofluorescence
than certified blank beads. Any suggestions on how to incorporate this in
the calculations of cell-bound FITC molecules on human RBCs?

Any suggestion appreciated ...

Best regards

Ulrik Sprogoe-Jakobsen

Dept. Clinical Immunology
Odense University Hospital
DK-5000 Odense C
Phone: +45 6541 3576
Fax: +45 6612 7975

• Next message: Frederic Preffer: "Low level fluorescence"
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CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories and distributed free of charge as an educational service to the cytometry community. If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL, Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu EMAIL robinson@flowcyt.cyto.purdue.edu