Synthetic polymer particles for use as standards and calibrators in flow cytometry

A method for calibrating or standardizing a flow cytometry instrument for particle analysis using substantially spherical synthetic particles as calibrators or standards, said calibration or standard synthetic particles having predetermined volume and refractive index values, wherein said instrument provides at least one light scatter-related signal from particles passing through said instrument for analysis, comprising:

• a) passing said calibration or standard synthetic particles, having average particle diameters and refractive indices essentially reproducible of particles to be assayed in a liquid flow stream, essentially one cell at a time through an incident beam of light;
• b) detecting a first measurable light scatter signal from said calibration or standard synthetic particles passing through said beam of light;
• c) detecting a second measurable signal from said calibration or standard synthetic particles;
• d) determining the intensity of said first light scatter signal and said second signal, thereby producing a pair of signal intensities;
• e) converting said pair of signal intensities into volume and refractive index values over a range of pairs of signals to provide standardized signal-gains to the flow cytometer;
• and
• f) calibrating the flow cytometer for i) volume and refractive index measurements and for ii) counting analyzable particles having signals within appropriate volume and refractive index ranges.

A method for calibrating or standardizing a flow cytometry instrument for particle analysis using substantially spherical synthetic particles as calibrators or standards, said calibration or standard synthetic particles having characteristic and known volume and refractive index values, wherein said instrument provides at least one light scatter-related signal from particles passing through said instrument for analysis, comprising:

• a) passing said calibration or standard synthetic particles, having average particle diameters and refractive indices essentially reproducible of particles to be assayed in a liquid flow stream, essentially one cell at a time through an incident beam of light;
• b) detecting first and second measurable light scatter signals from said calibration or standard synthetic particles passing through said beam of light;
• c) determining the intensities of said first and second light scatter signals, thereby producing a pair of signal scattering intensities;
• d) converting said pair of signal scattering intensities into volume and refractive index values over a range of pairs of signals to provide standardized signal-gains to the flow cytometer;
• and
• e) calibrating the flow cytometer for i) volume and refractive index measurements and for ii) counting analyzable particles having signals within appropriate volume and refractive index ranges.