STANDARDIZING FLOW-CYTOMETRY - CONSTRUCTION OF A STANDARDIZED FLUORESCENCE CALIBRATION PLOT USING MATCHING SPECTRAL CALIBRATORS

Calibration of flow cytometers is becoming an increasingly important i ssue for both quality control of instrument performance and quantitati on of antibody binding capacity of cells, Due to the numerous differen t instruments and analysis software currently available, a standardize d method of calibration is necessary if interlaboratory comparison of instrument performance and antibody binding is to be achieved, This re port describes a new methodology to obtain a standard calibration plot that can be derived from all instruments and from which specific inst rument-independent performance parameters may be calculated that can b e used to directly compare the performance and setup of these instrume nts, The requirements that the calibrated standards must meet are disc ussed, as well as the acceptable ranges proposed for the instrument-in dependent performance parameters, In addition, data are presented from standard calibration plots generated by different flow cytometers in numerous laboratories. The corresponding Primary Performance Parameter s calculated from these plots are presented and compared, It is expect ed that the use of this calibration method may help standardize flow c ytometric measurements and will provide instrument-independent perform ance parameters to monitor quality control of instruments and reagents . (C) 1996 Wiley-Liss, Inc.