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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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LED Thermo Flow &#8212; Combining Optogenetics with Flow Cytometry
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Practical time-gated luminescence flow cytometry. I: concepts.

Dayong Jin1, Russell Connally, James Piper

  • 1Centre for Lasers and Applications, Division of Information and Communication Sciences, Macquarie University, NSW, Australia. jin@ics.mq.edu.au

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|September 18, 2007
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Summary

Time-gated luminescence (TGL) flow cytometry enables high-speed cell analysis by suppressing background noise. New UV LED technology and practical models achieve 100% detection efficiency for labeled microorganisms.

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Optical Physics

Background:

  • Time-gated detection effectively reduces autofluorescence and light scatter in luminescence-based assays.
  • Applying time-gated detection to flow cytometry presents challenges in achieving high throughput for rapidly flowing samples.
  • Existing methods struggle to balance high cell analysis rates with efficient detection in flow cytometry.

Purpose of the Study:

  • To develop practical methods for high-throughput cell analysis using time-gated luminescence (TGL) flow cytometry.
  • To investigate the feasibility of new-generation UV LEDs as excitation sources for TGL flow cytometry.
  • To analyze the theoretical requirements and practical implementation of TGL in a continuous flow system.

Main Methods:

  • Investigated spatial effects of long-lived luminescence in fast-flowing samples.
  • Theoretically analyzed excitation and detection parameters for TGL flow cytometry.
  • Developed and evaluated two models: a triggered mode and a continuous flow-section mode.
  • Modeled a system using UV LED excitation for europium dye-labeled targets.

Main Results:

  • New-generation UV LEDs are suitable excitation sources for TGL flow cytometry.
  • The triggered model operates at low repetition rates, while the flow-section model allows high cell arrival rates.
  • Demonstrated feasible detection of europium dye-labeled beads with a signal-to-background ratio up to 11:1.
  • Achieved 100% detection efficiency at high cell analysis rates.

Conclusions:

  • Time-gated luminescence flow cytometry, particularly with UV LED excitation, offers a practical solution for high-throughput cell analysis.
  • The developed flow-section model enables significantly higher cell analysis rates compared to triggered systems.
  • This approach effectively suppresses background noise, enhancing signal detection for luminescence-labeled targets.