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13 March 2024 Autofluorescence lifetime flow cytometry with time-correlated single-photon counting
Kayvan Samimi, Ojaswi Pasachhe, Emmanuel Contreras Guzman, Amani A. Gillette, Dan L. Pham, Kathy J. Wiech, Jeremiah Riendeau, Melissa C. Skala
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Proceedings Volume PC12853, High-Speed Biomedical Imaging and Spectroscopy IX; PC1285307 (2024) https://doi.org/10.1117/12.3003561
Event: SPIE BiOS, 2024, San Francisco, California, United States
Abstract
Autofluorescence lifetime imaging microscopy is sensitive to functional or treatment-induced metabolic changes in single cells. However, this label-free technique typically relies on laser-scanning microscopes that are not common for single-cell assessments in immunology. Here, we present a microfluidic lifetime-sensitive flow cytometer that measures time-domain decays of the metabolic co-enzyme NAD(P)H from single cells with picosecond temporal resolution using time-correlated single-photon counting (TCSPC) at a throughput of tens of cells per second. The system uses a picosecond-pulsed ultraviolet diode laser, alkali photomultiplier tubes, an FPGA-based time tagger, and provides real-time phasor-based classification (i.e., gating) of flowing cells for applications in immunology.
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Kayvan Samimi, Ojaswi Pasachhe, Emmanuel Contreras Guzman, Amani A. Gillette, Dan L. Pham, Kathy J. Wiech, Jeremiah Riendeau, and Melissa C. Skala "Autofluorescence lifetime flow cytometry with time-correlated single-photon counting", Proc. SPIE PC12853, High-Speed Biomedical Imaging and Spectroscopy IX, PC1285307 (13 March 2024); https://doi.org/10.1117/12.3003561
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KEYWORDS
Time correlated single photon counting
Autofluorescence
Flow cytometry
Equipment
Fluorescence lifetime imaging
Semiconductor lasers
Single photon detectors
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