Fluorescent-particle analyzer with timing alignment for analog p

Details Fluorescent-particle analyzer with timing alignment for analog p Fluorescent-particle analyzer with timing alignment for analog p

1995-04-06

1997-10-28

Tokar, Michael J.

Radiant energy

Luminophor irradiation

2504591, 2504621, 250226, 356 39, 356411, G01J 158, G01J 351, G01N 2164, G01N 3348

Patent

active

056820380

ABSTRACT:
A flow cytometry system includes a flow tube through which particles marked with different fluorochromes pass from a first location, illuminated by a red laser, to a second location, illuminated by a blue laser. A "red" photodetector is optically coupled to detect red fluorescence from both locations. "Yellow" and "green" photodetectors respectively detect yellow and green fluorescence from the second location, while a "scattered light" detector detects scattered light from the second location. During a sample run, the red photodetector can output pulses that correspond primarily to APC fluorochrome at the first location and to PerCP at the second location respectively. A delay device delays the APC pulse relative to the PerCP pulse so that the peaks can be scaled and subtracted in the analog electrical domain to remove APC/PerCP crosstalk. The delay is calibrated using a set of APC tagged cells. Each of these cells generates a red fluorescence electrical pulse while at the first location and a scattered light pulse while at the second location. The average time differential between these two pulses for the set of timing cells determines the target delay. This is compared with the actual delay imposed by the delay device. If the error is greater than 0.5 .mu.S, the imposed delay can be adjusted until the delay is below 0.5 .mu.S, or within the tolerance required for reliable difference pulses to be generated by the pulse subtractor.

REFERENCES:
patent: 4573796 (1986-03-01), Martin et al.
patent: 5150313 (1992-09-01), Engh et al.
patent: 5168164 (1992-12-01), Urakami et al.
patent: 5270548 (1993-12-01), Steinkamp
"Improved Multilaser/Multiparameter Flow Cytometer For Analysis And Sorting Of Cells And Particles," by John A. Steinkamp, et al., Rev. Sci. Instrum. 62 (11), Nov. 1991, pp. 2751-2764.
"Cytomat-R: A Computer-Controlled Multiple Laser Source Multiparameter Flow Cytophotometer System," Authors: Howard M. Shapiro, et al., The Journal of Histochemistry and Cytochemistry, vol. 25, No. 7, pp. 836-844, 1977.
"A Generalized Machine For Automated Flow Cytology System Design," Authors: Raul Curbelo, et al., The Journal of Histochemistry And Cytochemistry, vol. 24, No. 1, pp. 388-395, 1976.
"Internal Calibration To Absolute Values in Flowthrough Particle Size Analysis," Authors: W.G. Eisert, et al., Rev. Sci. Instrum., 49(12), Dec. 1978, pp. 1617-1621.
"Laser Flow Cytometric Light Scatter And Fluorescence Pulse Rise-Time Sizing of Mammalian Cells.sup.1," Authors: James F. Leary, et al., The Journal of Histochemistry and Cytochemistry, vol. 27, No. 1, pp. 315-320, 1979.
"A Real-Time Delay Monitor For Flow-System Cell Sorters.sup.1," Authors: J.C. Martin, et al., The Journal of Histochemistry and Cytochemistry, vol. 27, No. 1, pp. 277-279, 1979.

Affiliated with

Also associated with

Rating

Fluorescent-particle analyzer with timing alignment for analog p does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Fluorescent-particle analyzer with timing alignment for analog p, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Fluorescent-particle analyzer with timing alignment for analog p will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-1027881