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Proceedings Paper

UV LED excited time-gated luminescence flow cytometry: evaluation for rare-event particle counting
Author(s): Dayong Jin; Belinda Ferrari; Robert C. Leif; Sean Yang; Lidia M. Vallarino; John Williams; James Piper
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Paper Abstract

Flow cytometric detection of specific rare-event targets within high-background samples such as water or food are frequently defeated by the extremely large population of non-target background particles. Time-gated detection of long lifetime fluorescence (>10μs) labeled microbial targets has been proven highly efficient in suppressing this non-target autofluorescent (<0.1μs) background. A time-gated luminescence (TGL) flow cytometer using UV LED excitation has demonstrated the successful detection of rare-event particles in high autofluorescence background samples. In this report, high-quality 5μm europium beads were made (homogenous intensity and aggregation free) for a detailed evaluation of the prototype performance. The known number of beads (10±2, 100±20 and 1000±100) were first sorted by a conventional flow cytometry sorter, and spiked into an environmental water concentrate (1 ml; containing >10 million non-target particles). The recovery rate for counting these very-rare-event particles using the TGL flow cytometer was then found to be 100%±20% between bead concentrations evaluated.

Paper Details

Date Published: 29 February 2008
PDF: 11 pages
Proc. SPIE 6859, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VI, 68590O (29 February 2008); doi: 10.1117/12.762077
Show Author Affiliations
Dayong Jin, Macquarie Univ. (Australia)
Belinda Ferrari, Macquarie Univ. (Australia)
Robert C. Leif, Newport Instruments (United States)
Sean Yang, Newport Instruments (United States)
Lidia M. Vallarino, Virginia Commonwealth Univ. (United States)
John Williams, Virginia Commonwealth Univ. (United States)
James Piper, Macquarie Univ. (Australia)

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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