Multi-illumination-source flow particle analyzer with inter-location emissions crosstalk cancelation
First Claim
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1. A particle analyzer comprising:
- flow means for flowing particles, including a first particle type and a second particle type, through a first location and through a second location;
first illumination means for illuminating said particles at said first location with light having a first spectral distribution;
second illumination means for illuminating said particles at said second location with light having a second spectral distribution different from said first spectral distribution;
first detection means for generating a first indicator when a particle of said first particle type is at said first location;
second detection means for generating a second indicator when a particle of said second particle type is at said second location, said second detection means also providing a third indicator when a particle of said first particle type is at said first detection location; and
cancellation means for at least partially canceling said third indicator in response to generation of said first indicator.
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Abstract
A multi-laser flow-cytometry system provides for inter-location emissions crosstalk cancellation, through either pulse subtraction or gating, to avoid false triggering based on shadow (pre- and post-) pulses that occur when red-excited red emissions are detected by a photodetector arranged to detect blue-excited red emissions. This overcomes a problem of undercounting of counting beads that led to misdeterminations of cell counts when blue-excited red fluorescence detections were used as a trigger. The approach also provides for more accurate quantitative data regarding fluorochrome emissions amplitudes.
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Citations
13 Claims
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1. A particle analyzer comprising:
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flow means for flowing particles, including a first particle type and a second particle type, through a first location and through a second location; first illumination means for illuminating said particles at said first location with light having a first spectral distribution; second illumination means for illuminating said particles at said second location with light having a second spectral distribution different from said first spectral distribution; first detection means for generating a first indicator when a particle of said first particle type is at said first location; second detection means for generating a second indicator when a particle of said second particle type is at said second location, said second detection means also providing a third indicator when a particle of said first particle type is at said first detection location; and cancellation means for at least partially canceling said third indicator in response to generation of said first indicator. - View Dependent Claims (2, 3, 4, 5)
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6. A method of analyzing particles, including particles of a first type and particles of a second type, said method comprising the steps of:
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flowing said particles through a first detection location illuminated by light of a first spectral distribution and through a second detection location illuminated by light of a second spectral distribution different from said first spectral distribution; generating a first indication when a particle of said first type is at said first detection location, generating a second indication along a second channel when a particle of said second type is at said second detection location, and generating a third indication along said second channel when said particle of said first type is at said first detection location; and at least partially canceling said third indicator in response to said first indicator. - View Dependent Claims (7, 8, 9, 10)
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11. A particle analyzer comprising:
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flow means for flowing particles, including a first particle, through a first location and through a second location; first illumination means for illuminating said particles at said first location; second illumination means for illuminating said particles at said second location; first detection means for generating a first indicator when said first particle is at said first location; second detection means for generating a second indicator when said first particle is at said second location; and cancellation means for at least partially canceling crosstalk in said first indicator and generated when said first particle is at said second location.
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12. A particle analyzer comprising:
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flow means for flowing particles through a first location and through a second location; first illumination means for illuminating said first location with light having a first source spectral distribution; second illumination means for illuminating said second location with light having a second source spectral distribution different from said first source spectral distribution; first detection means for generating a first indicator when light of a first emissions spectral distribution is emitted from said first location; second detection means for generating a second indicator when light of a second emissions spectral distribution is emitted from said second location, said second detection means also providing a third indicator when light of a third emissions spectral distribution is emitted from said first location; and cancellation means for at least partially canceling said third indicator in response to generation of said first indicator.
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13. A method of analyzing particles, including a first particle, said method comprising the steps of:
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flowing said particles through a first illuminated detection location and through a second illuminated detection location; generating a first indication when said first particle is at said first detection location, generating a second indication along a second channel when said first particle is at said second detection location; and at least partially canceling crosstalk generated at said second location when said particle is at said first location.
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Specification