Quantitative Flow Cytometry: History, Practice, Theory, Consensus, Inter-Laboratory Variation, and Present Status
G E. Marti, V E. Zenger, R F. Vogt, Adolfas Gaigalas
In the most general sense, quantitative fluorescence flow cytometry (QFCM) is the calibration of fluorescence intensity in terms of some reproducible scale. The present work gives a brief overview of the history of QFCM and points out what is the present, achievable level of inter-laboratory agreement. Two different calibration techniques are emphasized. One is based on microbeads with pre-assigned values of MESF, and the other is based on beads with immobilized goat anti-mouse immunoglobulin. The hope is that the beads with goat anti-mouse immunoglobulin provide a measure of the antibody binding capacity (ABC). For a given method, the measured values were consistent between laboratories. The MESF technique was dependent on fluorophore(FITC vs PE) whereas the ABS technique was less sensitive to the type of fluorophore used to label the CD4 antibody. The development at NIST of a more rigorous MESF assignment procedure should help to decrease the sensitivity of MESF based assays on fluorophore type.
ABC, cytometry, MESF, quantitative
, Zenger, V.
, Vogt, R.
and Gaigalas, A.
Quantitative Flow Cytometry: History, Practice, Theory, Consensus, Inter-Laboratory Variation, and Present Status, Cytotherapy
(Accessed December 5, 2023)