FRET-FLIM microscopy

Masilamani Elangovan, Richard Day, Ammasi Periasamy

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Citation (Scopus)

Abstract

Visualizing and quantifying protein-protein interactions is a recent trend in biomedical imaging. The current advances in fluorescence microscopy coupled with the development of new fluorescent probes provide the tools to study protein interactions in living specimens. Spectral bleed-through or cross talk is a problem in one- and two-photon microscopy to recognize whether one is observing the sensitized emission or the bleed-through signals. In contrast, FLIM (fluorescence lifetime imaging microscopy) or lifetime measurements are independent of excitation intensity or fluorophore concentration. The combination of FLIM and FRET will provide high spatial (nanometer) and temporal (nanoseconds) resolution when compared to steady state FRET imaging. Importantly, spectral bleed-through is not an issue in FLIM imaging because only the donor fluorophore lifetime is measured. The presence of acceptor molecules w/thin the local environment of the donor that permit energy transfer will influence the fluorescence lifetime of the donor. By measuring the donor lifetime in the presence and the absence of acceptor one can accurately calculate the FRET efficiency and the distance between donor- and acceptor-labeled proteins. Moreover, the FRET-FLIM technique allows monitoring more than one pair of protein interactions in a single living cell.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
EditorsA. Periasamy, P.T.C. So
Pages267-273
Number of pages7
Volume4620
DOIs
StatePublished - 2002
Externally publishedYes
EventMultiphoton Microscopy in the Biomedical Sciences II - San Jose, CA, United States
Duration: Jan 20 2002Jan 22 2002

Other

OtherMultiphoton Microscopy in the Biomedical Sciences II
CountryUnited States
CitySan Jose, CA
Period1/20/021/22/02

Fingerprint

Microscopic examination
Fluorescence
microscopy
Imaging techniques
life (durability)
fluorescence
Proteins
proteins
Fluorophores
Photons
Fluorescence microscopy
photons
interactions
Energy transfer
temporal resolution
Cells
energy transfer
Molecules
Monitoring
trends

Keywords

  • Fluorescence lifetime imaging (FLIM)
  • Fluorescence resonance energy transfer (FRET)
  • Protein-protein interactions
  • Two-photon excitation

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

Cite this

Elangovan, M., Day, R., & Periasamy, A. (2002). FRET-FLIM microscopy. In A. Periasamy, & P. T. C. So (Eds.), Proceedings of SPIE - The International Society for Optical Engineering (Vol. 4620, pp. 267-273) https://doi.org/10.1117/12.470682

FRET-FLIM microscopy. / Elangovan, Masilamani; Day, Richard; Periasamy, Ammasi.

Proceedings of SPIE - The International Society for Optical Engineering. ed. / A. Periasamy; P.T.C. So. Vol. 4620 2002. p. 267-273.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Elangovan, M, Day, R & Periasamy, A 2002, FRET-FLIM microscopy. in A Periasamy & PTC So (eds), Proceedings of SPIE - The International Society for Optical Engineering. vol. 4620, pp. 267-273, Multiphoton Microscopy in the Biomedical Sciences II, San Jose, CA, United States, 1/20/02. https://doi.org/10.1117/12.470682
Elangovan M, Day R, Periasamy A. FRET-FLIM microscopy. In Periasamy A, So PTC, editors, Proceedings of SPIE - The International Society for Optical Engineering. Vol. 4620. 2002. p. 267-273 https://doi.org/10.1117/12.470682
Elangovan, Masilamani ; Day, Richard ; Periasamy, Ammasi. / FRET-FLIM microscopy. Proceedings of SPIE - The International Society for Optical Engineering. editor / A. Periasamy ; P.T.C. So. Vol. 4620 2002. pp. 267-273
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