UV LED excited time-gated luminescence flow cytometry: evaluation for rare-event particle counting

Dayong Jin; Belinda Ferrari; Robert C. Leif; Sean Yang; Lidia M. Vallarino; John Williams; James Piper

doi:10.1117/12.762077

29 February 2008 UV LED excited time-gated luminescence flow cytometry: evaluation for rare-event particle counting

Dayong Jin, Belinda Ferrari, Robert C. Leif, Sean Yang, Lidia M. Vallarino, John Williams, James Piper

Proceedings Volume 6859, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VI; 68590O (2008) https://doi.org/10.1117/12.762077
Event: SPIE BiOS, 2008, San Jose, California, United States

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.

Citation Download Citation

Dayong Jin, Belinda Ferrari, Robert C. Leif, Sean Yang, Lidia M. Vallarino, John Williams, and James Piper "UV LED excited time-gated luminescence flow cytometry: evaluation for rare-event particle counting", Proc. SPIE 6859, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VI, 68590O (29 February 2008); https://doi.org/10.1117/12.762077

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