Fluorescence Lifetime Imaging

Klaus Suhling*, Liisa Hirvonen, James Levitt, Pei-Hua Chung, Carolyn Tregidgo, Dmitri A. Rusakov, Kaiyu Zheng, Simon Ameer-Beg, Simon Poland, Simao Coelho Pereira Coelho, Robert Henderson, Nikola Krstajic

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

6 Citations (Scopus)


Fluorescence lifetime imaging (FLIM) is a key fluorescence microscopy technique to map the environment and interaction of fluorescent probes. It can report on photophysical events that are difficult or impossible to observe by fluorescence intensity imaging, because FLIM is largely independent of the local fluorophore concentration and excitation intensity. Many FLIM applications relevant for biology concern the identification of Förster resonance energy transfer (FRET) to study protein interactions and conformational changes. In addition, FLIM has been used to image viscosity, temperature, pH, refractive index, and ion and oxygen concentrations, all at the cellular level. The basic principles and recent advances in the application of FLIM, FLIM instrumentation, molecular probe, and FLIM detector development will be discussed.

Original languageEnglish
Title of host publicationHandbook of Photonics for Biomedical Engineering
PublisherSpringer Netherlands
Number of pages53
ISBN (Electronic)9789400750524
ISBN (Print)9789400750517
Publication statusPublished - 29 Mar 2017


  • Anisotropy
  • Fluorescence anisotropy imaging (FAIM)
  • Fluorescence enhancement
  • Fluorescence microscopy
  • Fluorescence spectroscopy
  • Förster resonance energy transfer (FRET)
  • Plasmonics
  • Time-correlated single-photon counting (TCSPC)
  • Time-resolved fluorescence anisotropy imaging (TR-FAIM)
  • Total internal reflection fluorescence (TIRF)


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