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Optics Express

Optics Express

  • Editor: Michael Duncan
  • Vol. 14, Iss. 25 — Dec. 11, 2006
  • pp: 12217–12229

FRET-FLIM at nanometer spectral resolution from living cells

Deepak K. Nair, Mini Jose, Thomas Kuner, Werner Zuschratter, and Roland Hartig  »View Author Affiliations

Optics Express, Vol. 14, Issue 25, pp. 12217-12229 (2006)

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We report the investigation of Foerster’s Resonance Energy Transfer dynamics in GFP based tandem constructs in living T-cells using a combination of Fluorescence Lifetime Imaging Microscopy (FLIM) and Fluorescence Lifetime Micro-Spectroscopy (FLMS) at picosecond time resolution and nanometer spectral resolution. The involvement of multiple lifetimes of CFP in energy transfer was analyzed by plotting pre-exponential factors of individual lifetimes along the wavelength resulting in the Decay Associated Spectra (DAS). A change in the amplitude of pre-exponential factors from positive to negative at the acceptor emission maxima was used as a confirmation of FRET in the multiexponential lifetime analysis.

© 2006 Optical Society of America

OCIS Codes
(170.6920) Medical optics and biotechnology : Time-resolved imaging
(180.1790) Microscopy : Confocal microscopy
(320.4240) Ultrafast optics : Nanosecond phenomena

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: September 20, 2006
Revised Manuscript: October 30, 2006
Manuscript Accepted: October 30, 2006
Published: December 11, 2006

Virtual Issues
Vol. 2, Iss. 1 Virtual Journal for Biomedical Optics

Deepak K. Nair, Mini Jose, Thomas Kuner, Werner Zuschratter, and Roland Hartig, "FRET-FLIM at nanometer spectral resolution from living cells," Opt. Express 14, 12217-12229 (2006)

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