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Related Experiment Video

Updated: Nov 21, 2025

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
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Published on: February 9, 2012

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High-speed compressed-sensing fluorescence lifetime imaging microscopy of live cells.

Yayao Ma1,2, Youngjae Lee1,3, Catherine Best-Popescu1,3

  • 1Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801.

Proceedings of the National Academy of Sciences of the United States of America
|January 12, 2021
PubMed
Summary

We developed compressed fluorescence lifetime imaging microscopy (FLIM) for high-speed live-cell imaging. This technique captures detailed cellular dynamics in a single exposure, reducing motion artifacts and phototoxicity.

Keywords:
FLIMcompressed imaginghigh-speed

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Area of Science:

  • Microscopy
  • Biophysics
  • Cell Biology

Background:

  • Fluorescence Lifetime Imaging Microscopy (FLIM) is crucial for studying cellular processes.
  • Traditional FLIM methods face limitations in speed, motion artifacts, and phototoxicity.
  • High-speed, high-resolution live-cell imaging remains a significant challenge.

Purpose of the Study:

  • To introduce a novel compressed sensing-based FLIM system for high-speed live-cell imaging.
  • To overcome the limitations of conventional FLIM techniques.
  • To enable the study of transient biological dynamics at the microscopic level.

Main Methods:

  • Developed a compressed sensing scheme for FLIM.
  • Simultaneously recorded time-lapse fluorescence decay using pulsed laser excitation.
  • Acquired widefield fluorescence lifetime images within a single camera exposure.
  • Achieved imaging speeds up to 100 Hz.

Main Results:

  • Demonstrated high-resolution, high-speed FLIM of live cells.
  • The compressed FLIM system acquired images in a single exposure, eliminating motion artifacts.
  • Minimized photobleaching and phototoxicity compared to traditional methods.
  • Successfully imaged various transient dynamics at the microscopic scale.

Conclusions:

  • Compressed FLIM offers a powerful new tool for live-cell imaging.
  • The technique significantly enhances imaging speed and reduces artifacts.
  • Enables advanced studies of dynamic biological processes with minimal sample damage.