Abstract: Methods of assessing the suitability of a fluorochrome panel for use in a flow cytometric protocol analyzing a biological sample are provided. Methods of interest include, with a processor, receiving an initial fluorochrome panel, a plurality of population identifiers each referring to a particle population, and an instrument identifier. Methods additionally include, creating a set of population-marker pairs, generating a set of separability metrics each predicting a measure of statistical distance between particle populations in flow cytometer data space, aggregating the set of separability metrics into a panel score, and evaluating the panel score. Systems and non-transitory computer readable storage media for assessing the suitability of a fluorochrome panel for use in a flow cytometric protocol analyzing a biological sample are also provided.

Abstract: Methods of determining an ideal gain for a detector in a light detection system of a flow cytometer are provided. Methods of interest include irradiating a flow stream with light from a light source, incrementally increasing the gain of the detector such that the detector collects light from the flow stream at each of a plurality of successively increasing gains, obtaining a baseline noise level from the detector at each gain in the plurality of successively increasing gains, calculating a limit of detection (LoD) for each gain in the plurality of successively increasing gains, and assessing the calculated LoDs to determine the ideal gain. Systems and computer readable storage media for practicing the invention are also provided.

Abstract: Aspects of the present disclosure include methods for determining photodetector gain for a plurality of photodetectors in a light detection system. Methods according to certain embodiments include applying a reference voltage to each photodetector in the light detection system, generating a reference data signal for each photodetector at the reference voltage, irradiating with a light source the photodetectors at a plurality of different applied voltages, generating output data signals for each photodetector at each of the plurality of different voltages and calculating gain of the photodetectors at each of the plurality of different applied voltages based on the output data signals for each photodetector at each applied voltage and the reference data signal. Systems (e.g., particle analyzers) having a light source and a light detection system that includes a plurality of photodetectors for practicing the subject methods are also described. Non-transitory computer readable storage medium are also provided.