Multi-spectral filter profiling and quality control for flow cytometry
First Claim
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1. A system comprising:
- a flow cytometer including;
at least one sample illumination source, wherein each sample illumination source is configured to deliver light to a corresponding sample location, thereby causing sample light to be emitted by or scattered off of particles in the corresponding sample location;
focusing optics that are configured to direct the sample light from each sample location along one or more optical paths, wherein each optical path passes through a corresponding one or more optical filter elements and terminates at a corresponding detector that is configured to produce output data indicative of the measured intensity of light reaching that detector;
a calibration light source that is configured to independently emit different spectral profiles of calibration light at different times, each spectral profile of calibration light having one or more peaks at different wavelengths, such that the emitted calibration light is directed along at least a portion of each of the optical paths, wherein the calibration light source is different than the sample illumination source;
one or more processors; and
a memory that stores computer-executable instructions that, when executed by the one or more processors, cause the one or more processors to;
receive the output data from each of the detectors responsive to that detector receiving calibration light from the calibration light source, anddetermine filtering characteristics for the optical filter elements along each optical path by analyzing the output data produced during the emission of at least two different spectral profiles of calibration light by the calibration light source.
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Abstract
Disclosed is a system and method for characterizing optical filters in a flow cytometer and optionally checking the operation of detectors in the flow cytometer. In some embodiments, the system may utilize an LED board having an opening through which the fluorescence and side scatter beams, rays, or images pass and light emitting diodes around the opening that emit light having different spectral profiles. The different spectral profiles allow the system to identify the optical filters that are placed in the flow cytometer, to verify detector operation, to assist in instrumentation troubleshooting, and to provide a quantitative reference for detector comparison.
19 Citations
26 Claims
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1. A system comprising:
a flow cytometer including; at least one sample illumination source, wherein each sample illumination source is configured to deliver light to a corresponding sample location, thereby causing sample light to be emitted by or scattered off of particles in the corresponding sample location; focusing optics that are configured to direct the sample light from each sample location along one or more optical paths, wherein each optical path passes through a corresponding one or more optical filter elements and terminates at a corresponding detector that is configured to produce output data indicative of the measured intensity of light reaching that detector; a calibration light source that is configured to independently emit different spectral profiles of calibration light at different times, each spectral profile of calibration light having one or more peaks at different wavelengths, such that the emitted calibration light is directed along at least a portion of each of the optical paths, wherein the calibration light source is different than the sample illumination source; one or more processors; and a memory that stores computer-executable instructions that, when executed by the one or more processors, cause the one or more processors to; receive the output data from each of the detectors responsive to that detector receiving calibration light from the calibration light source, and determine filtering characteristics for the optical filter elements along each optical path by analyzing the output data produced during the emission of at least two different spectral profiles of calibration light by the calibration light source. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. A method for determining filtering characteristics for a plurality of optical filter elements in a flow cytometry system, wherein a different subset of a plurality of optical paths passes through each of the optical filter elements, and wherein each optical path directs emitted or scattered sample light from a sample location to a corresponding detector, the method comprising:
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emitting different spectral profiles of calibration light from a calibration light source of the flow cytometry system at different times, wherein each spectral profile of calibration light has one or more intensity peaks at different wavelengths, and wherein the calibration light source is different than a sample illumination source of the flow cytometry system, the sample illumination source being configured to deliver light to each of the sample locations, thereby producing the emitted or scattered sample light; directing some of the calibration light along at least a portion of each of the optical paths; measuring, for each different spectral profile of calibration light, the light intensity at each of the detectors, wherein each of the detectors produces output data that is indicative of the measured light intensity of the calibration light that reaches the detector; and analyzing the output data from one of the detectors produced during the emission of at least two different spectral profiles of calibration light by the calibration light source to determine the filtering characteristics of the optical filter elements along the optical path corresponding to that detector. - View Dependent Claims (19, 20, 21, 22, 23, 24, 25, 26)
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Specification