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Related Experiment Video

Updated: Jun 25, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
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Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

Analog mean-delay method for high-speed fluorescence lifetime measurement.

Sucbei Moon1, Youngjae Won, Dug Young Kim

  • 1Department of Information and Communications, Gwangju Institute of Science and Technology, Buk-gu, Gwangju, Republic of Korea 500-712. moonsb@gist.ac.kr

Optics Express
|February 17, 2009
PubMed
Summary

We developed a new high-speed fluorescence lifetime measurement technique using the analog mean-delay (AMD) method. This approach enhances speed and accuracy for time-resolved spectroscopy and FLIM without photon-rate limits.

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Area of Science:

  • Spectroscopy and Microscopy
  • Photonics and Optical Science

Background:

  • Fluorescence lifetime measurements are crucial for time-resolved spectroscopy and FLIM.
  • Existing methods often face limitations in speed and photon detection rates.

Purpose of the Study:

  • To introduce a novel high-speed fluorescence lifetime measurement scheme.
  • To evaluate the performance of the analog mean-delay (AMD) method for speed, accuracy, and precision.

Main Methods:

  • Acquisition of time-domain fluorescence decay signals as analog waveforms.
  • Extraction of lifetime information from the mean temporal delay of the analog signal.
  • Utilizing simultaneous multi-photon detection, bypassing single-photon counting limitations.

Main Results:

  • Demonstrated high-speed measurements with photon detection rates up to 10^8 photons/second.
  • Achieved a fluorescence lifetime accuracy of 3.2 ns with a 3% standard deviation.
  • Obtained a measurement rate of 56,300 lifetime determinations per second with near shot-noise-limited photon economy.

Conclusions:

  • The AMD method offers excellent measurement performance in accuracy, precision, and speed.
  • This technique enables significantly faster fluorescence lifetime determination.
  • The method is suitable for dynamical time-resolved spectroscopy and high-speed FLIM applications.