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Luminescence lifetime encoding in time-domain flow cytometry.

Daniel Kage1,2, Katrin Hoffmann1, Marc Wittkamp3

  • 1Federal Institute for Materials Research and Testing (BAM), Biophotonics Division 1.2, Richard-Willstätter-Str. 11, D-12489, Berlin, Germany.

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Time-resolved flow cytometry offers a new bioanalytical approach using lifetime-encoded beads. This study details a compact platform for straightforward time-domain measurements, enhancing multiplexing capabilities.

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Area of Science:

  • Bioanalytical chemistry
  • Biophysics
  • Analytical instrumentation

Background:

  • Traditional spectral or intensity multiplexing in bioanalytics has limitations.
  • Phase-domain techniques dominate current time-resolved flow cytometry research.
  • A need exists for alternative, accessible time-resolved methods.

Purpose of the Study:

  • To present a novel flow cytometry platform utilizing time-resolved detection.
  • To demonstrate straightforward time-domain measurements with lifetime-encoded beads.
  • To assess the potential and limitations of this time-domain approach.

Main Methods:

  • Development of a compact flow cytometry setup for time-resolved measurements.
  • Utilizing beads with fluorescence lifetimes in the nanosecond range for encoding.
  • Performing general assessment, numerical calculations, and data analysis.

Main Results:

  • Successful implementation of a time-domain flow cytometry platform.
  • Demonstration of lifetime-encoded beads for multiplexing.
  • Analysis of resolution limits, data processing, and dye requirements.

Conclusions:

  • Time-domain flow cytometry offers a viable alternative to conventional multiplexing techniques.
  • The presented compact platform provides a straightforward approach to time-resolved bioanalysis.
  • Further research can optimize dye selection and data analysis for enhanced performance.