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Proceedings Paper

A comparison of avalanche photodiode and photomultiplier tube detectors for flow cytometry
Author(s): William G. Lawrence; Gyula Varadi; Gerald Entine; Edward Podniesinski; Paul K. Wallace
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Paper Abstract

Commercial flow cytometers use photomultiplier tubes (PMTs) for fluorescence detection. These detectors have high linear gain and broad dynamic range, but have limited sensitivity in the red and near infrared spectral regions. We present a comparison of avalanche photodiodes (APDs) and PMTs as detectors in flow cytometry instruments, and demonstrate improved sensitivity and resolution in the red and near infrared spectral regions using the APD. The relative performance of the PMT and APD were evaluated by simultaneously measuring the mean fluorescence intensity and coefficient of variation for emission from light emitting diode pulses, flow cytometry test beads, and fluorescently labeled cells. The relative signal to noise performance of the APD and PMT was evaluated over the 500 nm to 1050 nm wavelength range using pulsed light emitting diode light sources. While APDs have higher quantum efficiency but lower internal gain than PMTs, with appropriate external amplification the APD has signal to noise response that is comparable to PMTs in the 500 nm to 650 nm range and improved response in the 650 nm to 850 nm range The data demonstrates that the APD had performance comparable to the PMT in the spectral region between 500 to 650 nm and improved performance in the range of 650 to 1000 nm, where the PMT performance is quite poor. CD4 positive lymphocyte populations were easily identified in normal human blood both by APD and PMT using phycoerythrin labeled antibodies. In contrast, only the APD detector could resolve CD4 positive populations using 800 nm Quantum dot labeled antibodies.

Paper Details

Date Published: 29 February 2008
PDF: 11 pages
Proc. SPIE 6859, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VI, 68590M (29 February 2008); doi: 10.1117/12.758958
Show Author Affiliations
William G. Lawrence, Radiation Monitoring Devices, Inc. (United States)
Gyula Varadi, Radiation Monitoring Devices, Inc. (United States)
Gerald Entine, Radiation Monitoring Devices, Inc. (United States)
Edward Podniesinski, Roswell Park Cancer Institute (United States)
Paul K. Wallace, Roswell Park Cancer Institute (United States)


Published in SPIE Proceedings Vol. 6859:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VI
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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