, Volume 251, Issue 2, pp 293–305

Fluorescence lifetime imaging microscopy in the medical sciences

Special Issue: New/Emerging Techniques in Biological Microscopy

DOI: 10.1007/s00709-013-0598-4

Cite this article as:
Ebrecht, R., Don Paul, C. & Wouters, F.S. Protoplasma (2014) 251: 293. doi:10.1007/s00709-013-0598-4


The steady improvement in the imaging of cellular processes in living tissue over the last 10–15 years through the use of various fluorophores including organic dyes, fluorescent proteins and quantum dots, has made observing biological events common practice. Advances in imaging and recording technology have made it possible to exploit a fluorophore's fluorescence lifetime. The fluorescence lifetime is an intrinsic parameter that is unique for each fluorophore, and that is highly sensitive to its immediate environment and/or the photophysical coupling to other fluorophores by the phenomenon Förster resonance energy transfer (FRET). The fluorescence lifetime has become an important tool in the construction of optical bioassays for various cellular activities and reactions. The measurement of the fluorescence lifetime is possible in two formats; time domain or frequency domain, each with their own advantages. Fluorescence lifetime imaging applications have now progressed to a state where, besides their utility in cell biological research, they can be employed as clinical diagnostic tools. This review highlights the multitude of fluorophores, techniques and clinical applications that make use of fluorescence lifetime imaging microscopy (FLIM).


Organic dyes Quantum dots (QDs) Fluorescent proteins (FPs) Quenchers Förster resonance energy transfer (FRET) Fluorescence lifetime imaging microscopy (FLIM) Time-domain FLIM (TD FLIM) Frequency-domain FLIM (FD FLIM) Endogenous fluorophores 

Copyright information

© Springer-Verlag Wien 2014

Authors and Affiliations

  • René Ebrecht
    • 1
    • 2
  • Craig Don Paul
    • 1
    • 2
  • Fred S. Wouters
    • 1
    • 2
  1. 1.Department of Neuro- and Sensory PhysiologyUniversity Medicine GöttingenGöttingenGermany
  2. 2.Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB)GöttingenGermany

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