Flow cytometry apparatus and method
First Claim
1. Apparatus for identifying particles in a sample stream moving through a flow zone at a rate relative to the flow zone, the sample stream containing target particles, the apparatus comprising:
- means for illuminating the sample stream within the flow zone;
a detector for detecting light emitted or scattered from illuminated target particles within the flow zone;
a time delayed integration element for integrating the detected light, the time delayed integration synchronized to the sample stream flow rate; and
means for identifying target particles responsive to the time delayed integration element.
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Abstract
A flow cytometer for detecting target particles such as microorganisms including biological cells and viruses as well as molecular species. The flow cytometer includes a detection system involving a CCD having a time delay integration capability to thereby increase the signal from the target particle and decrease the noise detected by the CCD. Calibration particles can be included in the sample stream of the flow cytometer for coordinating the readout of the CCD with the rate of flow of the sample stream to improve the detection capability of the CCD. Statistical analysis techniques can also be used to determine the rate of flow of target particles in the sample stream to thereby coordinate the readout rate of the CCD with the rate of flow of the target particles.
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Citations
15 Claims
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1. Apparatus for identifying particles in a sample stream moving through a flow zone at a rate relative to the flow zone, the sample stream containing target particles, the apparatus comprising:
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means for illuminating the sample stream within the flow zone;
a detector for detecting light emitted or scattered from illuminated target particles within the flow zone;
a time delayed integration element for integrating the detected light, the time delayed integration synchronized to the sample stream flow rate; and
means for identifying target particles responsive to the time delayed integration element. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
a calibration particle in the sample flow; and
means for measuring the velocity of the calibration particle.
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9. The apparatus of claim 8, wherein the sample stream rate is measured in real time while target particles are being detected.
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10. The apparatus of claim 8 wherein the calibration particle is a target particle.
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11. The apparatus of claim 10, wherein the sample stream rate is measured in real time while target particles are being detected.
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12. The apparatus of claim 1, further including means for measuring the sample stream flow rate including:
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means for applying statistical analysis to data collected by pixels of the CCD to determine pixels related to a single target particle; and
means for measuring velocity of at least one target particle.
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13. The apparatus of claim 1, further including means for synchronizing the time delayed integration element and the sample stream flow rate by adjusting the time delayed integration element until target particles appear substantially as dots.
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14. The apparatus of claim 1, further including means for detecting two different target particle species, wherein each species produces a unique frequency or phase shift in its scattered or emitted light, wherein the detector includes means for differentiating frequency of phase shift.
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15. The apparatus of claim 1, further including a notch filter between the sample stream and the detector for filtering out undesired light.
Specification