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Emerging trends of fluorescence lifetime imaging microscopy (FLIM): advances, challenges, and prospects

12.29.25 | Higher Education Press

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Image:
FLIM principles. Schematic overview of fluorescence lifetime data acquisition and analysis in time−domain (td) and frequency−domain (fd) modes. In tdFLIM, photon arrival times are recorded after each pulsed excitation to build a decay histogram, which can be analyzed by multi-exponential curve fitting or transformed into the phasor space for model-free lifetime extraction. In fdFLIM, the emission is measured under high-frequency intensity modulation, yielding phase shift and demodulation factors Credit: HIGHER EDUCATON PRESS

This mini-review provides a concise overview of the latest advances in fluorescence lifetime imaging microscopy (FLIM). It discusses both time-domain and frequency-domain techniques, analysis methods − including phasor approaches and deep learning, and highlights applications in multiplexed imaging and quantitative biosensing. Furthermore, FLIM-empowered multimodal imaging approaches aimed at enhancing spatial and temporal resolution are discussed. Persistent challenges, including photon efficiency, probe sensitivity, and achieving high-speed imaging in live-cell environments, are critically assessed, outlining pathways toward future innovations.

10.52601/bpr.2025.250024

Experimental study

Not applicable

Emerging trends of fluorescence lifetime imaging microscopy (FLIM): advances, challenges, and prospects

1-Jan-2026

Keywords

Biophysics

Article Information

DOI
10.52601/bpr.2025.250024
Method of Research
Experimental study
Subject of Research
Not applicable
Article Publication Date
2026-01-01
Article Title
Emerging trends of fluorescence lifetime imaging microscopy (FLIM): advances, challenges, and prospects

Contact Information

Rong Xie
Higher Education Press
xierong@hep.com.cn

Source

Original Source
https://www.biophysics-reports.org/article/doi/10.52601/bpr.2025.250024

How to Cite This Article

APA:
Higher Education Press. (2025, December 29). Emerging trends of fluorescence lifetime imaging microscopy (FLIM): advances, challenges, and prospects. Brightsurf News. https://www.brightsurf.com/news/1EOWPW7L/emerging-trends-of-fluorescence-lifetime-imaging-microscopy-flim-advances-challenges-and-prospects.html
MLA:
"Emerging trends of fluorescence lifetime imaging microscopy (FLIM): advances, challenges, and prospects." Brightsurf News, Dec. 29 2025, https://www.brightsurf.com/news/1EOWPW7L/emerging-trends-of-fluorescence-lifetime-imaging-microscopy-flim-advances-challenges-and-prospects.html.