Fluorescence lifetime imaging (FLIM): Basic concepts and some recent developments

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

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    Abstract

    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 independent of the local fluorophore concentration and excitation intensity. One prominent FLIM application relevant for biological concerns is the identification of FRET to study protein interactions and conformational changes, but FLIM is also used to image viscosity, temperature, pH, refractive index and ion and oxygen concentrations, all at the cellular level, as well as cell and tissue autofluorescence. The basic principles and recent advances in the application of FLIM, FLIM instrumentation, data analysis, molecular probe and FLIM detector development will be discussed.

    Original languageEnglish
    Pages (from-to)3-40
    Number of pages38
    JournalMedical Photonics
    Volume27
    Early online date18 Mar 2015
    DOIs
    Publication statusPublished - May 2015

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    Keywords

    • Fluorescence microscopy
    • Fluorescence sensing
    • Fluorescence spectroscopy
    • Förster resonance energy transfer (FRET)
    • Time-correlated single photon counting (TCSPC)
    • Time-resolved fluorescence anisotropy imaging (TR-FAIM)

    Cite this

    Suhling, K., Hirvonen, L. M., Levitt, J. A., Chung, P. H., Tregidgo, C., Le Marois, A., Rusakov, D. A., Zheng, K., Ameer-Beg, S., Poland, S., Coelho, S., Henderson, R., & Krstajic, N. (2015). Fluorescence lifetime imaging (FLIM): Basic concepts and some recent developments. Medical Photonics, 27, 3-40. https://doi.org/10.1016/j.medpho.2014.12.001