UV LED excited time-gated luminescence flow cytometry: Concepts and experimental evaluation

Jin Dayong; Russell Connally; Jim Piper

doi:10.1117/12.685076

UV LED excited time-gated luminescence flow cytometry: Concepts and experimental evaluation

Jin Dayong^*, Russell Connally, Jim Piper

^*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceeding › Conference proceeding contribution › peer-review

2 Citations (Scopus)

Abstract

This paper presents experimental and theoretical studies of time-gated discrimination of long-lived luminescence (lifetime: 1-2000 μs) labelled target-organisms against non-target autofluorescence background (lifetime: <100 ns) in flow cytometry. A theoretical model of such a TGL flow cytometer is developed which takes account of flow speed, illumination and detection apertures, fluorescence label lifetime, and pulsed illumination and gated detection timing sequences. Ultraviolet LED and channel photomultiplier were found to be practical as pulsed excitation sources and gated detector for TGL flow cytometry. The prototype cytometer was constructed and optimized to operate at 6 k Hz repetition rate of TGL cycles consisting of 100 μs LED excitation and ∼60 μs gated detection. The spatial counting efficiency was evaluated by enumerating 5.5 μm diameter europium microspheres resulting in a counting accuracy approaching 100%.

Original language	English
Title of host publication	Smart Medical and Biomedical Sensor Technology IV
Editors	Brian M. Cullum, J. Chance Carter
Place of Publication	Bellingham, WA
Publisher	SPIE
Pages	1-8
Number of pages	8
Volume	6380
ISBN (Print)	0819464783, 9780819464781
DOIs	https://doi.org/10.1117/12.685076
Publication status	Published - 2006
Event	Smart Medical and Biomedical Sensor Technology IV - Boston, MA, United States Duration: 3 Oct 2006 → 4 Oct 2006

Other

Other	Smart Medical and Biomedical Sensor Technology IV
Country/Territory	United States
City	Boston, MA
Period	3/10/06 → 4/10/06

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10.1117/12.685076

Cite this

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title = "UV LED excited time-gated luminescence flow cytometry: Concepts and experimental evaluation",

abstract = "This paper presents experimental and theoretical studies of time-gated discrimination of long-lived luminescence (lifetime: 1-2000 μs) labelled target-organisms against non-target autofluorescence background (lifetime: <100 ns) in flow cytometry. A theoretical model of such a TGL flow cytometer is developed which takes account of flow speed, illumination and detection apertures, fluorescence label lifetime, and pulsed illumination and gated detection timing sequences. Ultraviolet LED and channel photomultiplier were found to be practical as pulsed excitation sources and gated detector for TGL flow cytometry. The prototype cytometer was constructed and optimized to operate at 6 k Hz repetition rate of TGL cycles consisting of 100 μs LED excitation and ∼60 μs gated detection. The spatial counting efficiency was evaluated by enumerating 5.5 μm diameter europium microspheres resulting in a counting accuracy approaching 100%.",

author = "Jin Dayong and Russell Connally and Jim Piper",

year = "2006",

doi = "10.1117/12.685076",

language = "English",

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editor = "Cullum, {Brian M.} and Carter, {J. Chance}",

booktitle = "Smart Medical and Biomedical Sensor Technology IV",

publisher = "SPIE",

address = "United States",

note = "Smart Medical and Biomedical Sensor Technology IV ; Conference date: 03-10-2006 Through 04-10-2006",

}

Dayong, J, Connally, R & Piper, J 2006, UV LED excited time-gated luminescence flow cytometry: Concepts and experimental evaluation. in BM Cullum & JC Carter (eds), Smart Medical and Biomedical Sensor Technology IV. vol. 6380, 63800L, SPIE, Bellingham, WA, pp. 1-8, Smart Medical and Biomedical Sensor Technology IV, Boston, MA, United States, 3/10/06. https://doi.org/10.1117/12.685076

UV LED excited time-gated luminescence flow cytometry: Concepts and experimental evaluation. / Dayong, Jin; Connally, Russell; Piper, Jim.
Smart Medical and Biomedical Sensor Technology IV. ed. / Brian M. Cullum; J. Chance Carter. Vol. 6380 Bellingham, WA: SPIE, 2006. p. 1-8 63800L.

Research output: Chapter in Book/Report/Conference proceeding › Conference proceeding contribution › peer-review

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N2 - This paper presents experimental and theoretical studies of time-gated discrimination of long-lived luminescence (lifetime: 1-2000 μs) labelled target-organisms against non-target autofluorescence background (lifetime: <100 ns) in flow cytometry. A theoretical model of such a TGL flow cytometer is developed which takes account of flow speed, illumination and detection apertures, fluorescence label lifetime, and pulsed illumination and gated detection timing sequences. Ultraviolet LED and channel photomultiplier were found to be practical as pulsed excitation sources and gated detector for TGL flow cytometry. The prototype cytometer was constructed and optimized to operate at 6 k Hz repetition rate of TGL cycles consisting of 100 μs LED excitation and ∼60 μs gated detection. The spatial counting efficiency was evaluated by enumerating 5.5 μm diameter europium microspheres resulting in a counting accuracy approaching 100%.

AB - This paper presents experimental and theoretical studies of time-gated discrimination of long-lived luminescence (lifetime: 1-2000 μs) labelled target-organisms against non-target autofluorescence background (lifetime: <100 ns) in flow cytometry. A theoretical model of such a TGL flow cytometer is developed which takes account of flow speed, illumination and detection apertures, fluorescence label lifetime, and pulsed illumination and gated detection timing sequences. Ultraviolet LED and channel photomultiplier were found to be practical as pulsed excitation sources and gated detector for TGL flow cytometry. The prototype cytometer was constructed and optimized to operate at 6 k Hz repetition rate of TGL cycles consisting of 100 μs LED excitation and ∼60 μs gated detection. The spatial counting efficiency was evaluated by enumerating 5.5 μm diameter europium microspheres resulting in a counting accuracy approaching 100%.

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UV LED excited time-gated luminescence flow cytometry: Concepts and experimental evaluation

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