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Updated: Jul 17, 2026
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Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
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
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.
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.
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