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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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

High-fidelity fast fluorescence lifetime imaging by event-based denoising.

Yiliang Zhou1,2,3, Yihong Xiao4, Jing Zhou1,2,3

  • 1Department of Automation, Tsinghua University, Beijing, China.

Nature Biotechnology
|July 15, 2026
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Summary

Event-based first-photon FLIM (EFLIM) significantly reduces photon requirements for fluorescence lifetime imaging microscopy. This breakthrough enables precise molecular measurements even in challenging deep-tissue and low-light conditions.

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Area of Science:

  • Biophotonics and advanced microscopy techniques.
  • Quantitative molecular imaging and analysis.

Background:

  • Fluorescence lifetime imaging microscopy (FLIM) offers valuable insights into molecular environments and interactions.
  • High photon counts limit FLIM's application in dynamic, deep-tissue imaging scenarios.

Purpose of the Study:

  • To develop a novel, low-light fluorescence lifetime imaging method.
  • To overcome the photon-starved limitations of conventional FLIM for in vivo applications.

Main Methods:

  • Introduced event-based first-photon FLIM (EFLIM), a self-supervised denoising approach.
  • Utilized a binary process representation for excitation events, reducing photon needs by over 100x.
  • Achieved accurate lifetime measurements with less than one photon per pixel.

Main Results:

  • Demonstrated robust fluorescence lifetime measurement at extremely low light levels.
  • Successfully observed transient intracellular dynamics and ligand-dependent molecular states.
  • Visualized vesicle-mediated lymphocyte contacts and human glioma tumor heterogeneity rapidly and label-free.

Conclusions:

  • EFLIM significantly enhances the applicability of FLIM for fast, deep-tissue, and low-light biological imaging.
  • The method shows strong potential for neuroscience, cell biology, immunology, and pathology research.
  • EFLIM enables in vivo probing of dynamic molecular processes with unprecedented sensitivity.