Methods for determining a photodetector gain correction factor for optical signals in a flow cytometer

A particle analyzer comprising:

• a light detection system comprising an imaging photodetector, wherein the light detection system is configured to:
• detect light from one or more laser beams irradiating across a horizontal axis of a flow stream perpendicular to the flow direction of the flow stream, the flow stream consisting of a single fluid column configured to move a single stream of particles across each laser beam along a common flow path;
• generate, for an imaging laser beam of the one or more laser beams, data signals in an imaging photodetector channel of the light detection system at a plurality of pixel locations, each pixel location corresponding to a different position across the horizontal axis of the flow stream irradiated by the one or more laser beams;
• and
• a processor comprising memory operably coupled to the processor wherein the memory comprises instructions stored thereon, which when executed by the processor, cause the processor to calculate a detector gain correction factor for each pixel location across the flow stream in response to the generated data signals, the detector gain correction factor comprising an adjustment to signal intensity of data signals generated in a photodetector channel for the pixel location such that intensity variation in the data signals of the photodetector channel across the flow stream is 10% or less.

A non-transitory computer readable storage medium comprising instructions stored thereon for determining a detector gain correction factor for application to flow cytometer data, the instructions comprising:

• algorithm for detecting light with a light detection system from one or more laser beams irradiating across a horizontal axis of a flow stream perpendicular to the flow direction of the flow stream, the flow stream consisting of a single fluid column configured to move a single stream of particles across each laser beam along a common flow path;
• algorithm for generating, for an imaging laser beam of the one or more laser beams, data signals in an imaging photodetector channel of the light detection system at a plurality of pixel locations, each pixel location corresponding to a different position across the horizontal axis of the flow stream irradiated by the one or more laser beams;
• and
• algorithm for calculating a detector gain correction factor for each pixel location across the flow stream in response to the generated data signals, the detector gain correction factor comprising an adjustment to signal intensity of data signals generated in a photodetector channel for the pixel location such that intensity variation in the data signals of the photodetector channel across the flow stream is 10% or less.