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MESFs were developed to provide a meaningful "absolute" unit of
fluorescence intensity which was instrument independent. MESFs for
standard particles are established via spectrofluormetry directly against
solutions of high purity fluorochome which have the same excitation and
emission spectra and are as responsive to the environment as the labeled
samples, e.g., pH. Meeting these criteria, automatically provides
correction for differences in extinction coefficients, quenching and
changes in quantum efficiency which would be necessary to consider if
actual numbers of fluorochrome molecules on the standard particles were
determined.
The following references may provide useful information:
Schwartz, A., Monograph: Fluorescent Microbead Standards, pub FCSC 1988.
Schwartz, A. and Fernandez-Repollet, E., Technical Aspects of Fluorescence
Quantitative Measurements by Flow Cytometry, Clinical Immunology
Newsletter vol 15 (6/7) pp. 73-77 1995.
Schwartz, A. and Fernendez-Repollet, E., Development of Clinical
Standards for Flow Cytometry, Annals of NY Acad. of Sci., vol 677 pp.
28-39, 1993.
Schwartz, A., Fernandez-Repollet, E., Vogt, R. and Gratama, J.W.,
Standardizing Flow Cytometry: Construction of a Standardized
Fluorescence Calibration Plot Using Matching Spectral Calibrators,
Cytometry (Comm in Clinical Cytometry) 26:22-31, 1996.
Note, the determining MESFs of a sample does not accurately indicate the
number of antibodies binding to the sample (Antibody Binding Capacity,
ABC) unless the "effective F/P" ratio or antibody binding standards are
used.
I hope this information clarifies what MESF units are and how they are
used.
Best regards,
Abe Schwartz, PhD
President, FCSC
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CD-ROM Vol 3 was produced by Monica M. Shively and other 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: (765)-494-0757;
FAX(765) 494-0517;
Web