FRET-FLIM at nanometer spectral resolution from living cells
Optics Express, Vol. 14, Issue 25, pp. 12217-12229 (2006)
http://dx.doi.org/10.1364/OE.14.012217
Enhanced HTML
Acrobat PDF (199 KB)
Abstract
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
History
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
Citation
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)
http://www.opticsinfobase.org/vjbo/abstract.cfm?URI=oe-14-25-12217
Sort: Year | Journal | Reset
References
- Y. Chen, J. D. Mills, and A. Periasamy, "Protein localization in living cells and tissues using FRET and FLIM," Differentiation 71, 528-541 (2003). [CrossRef] [PubMed]
- M. Elangovan, R. N. Day, and A. Periasamy, "Nanosecond fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy to localize the protein interactions in a single living cell," J. Microsc. 205, 3-14 (2002). [CrossRef] [PubMed]
- S. I. Murata, J. Kusba, G. Piszczek, I. Gryczynski, and J. R. Lakowicz, "Donor fluorescence decay analysis for energy transfer in double-helical DNA with various acceptor concentrations," Biopolymers 57, 306-315 (2000). [CrossRef] [PubMed]
- M. Tramier, I. Gautier, T. Piolot, S. Ravalet, K. Kemnitz, J. Coppey, C. Durieux, V. Mignotte, and M. Coppey-Moisan, "Picosecond-hetero-FRET microscopy to probe protein-protein interactions in live cells," Biophys. J. 83, 3570-3577 (2002). [CrossRef] [PubMed]
- J. W. Borst, M. A. Hink, A. V. Hoek, and A. J. W. G. Visser, "Effects of Refractive index and viscocity on Flourescence and Anisotropy Decays of Enhanced Cyan and Yellow Fluorescent Proteins," J. Fluoresc. 15, 153-160 (2005). [CrossRef] [PubMed]
- T. Kuner and G. J. Augustine, "A genetically encoded ratiometric indicator for chloride: capturing chloride transients in cultured hippocampal neurons," Neuron 27, 447-459 (2000). [CrossRef] [PubMed]
- M. Ormo, A. B. Cubitt, K. Kallio, L. A. Gross, R. Y. Tsien, and S. J. Remington, "Crystal structure of the Aequorea victoria green fluorescent protein," Science 273, 1392-1395 (1996). [CrossRef] [PubMed]
- J. R. Lakowicz, Principles of Fluorescence Spectroscopy, Second ed. (Kluwer Academic/Plenum Publishers, 1999).
- J. R. Knutson, D. G. Walbridge, and L. Brand, "Decay-associated fluorescence spectra and the heterogeneous emission of alcohol dehydrogenase," Biochemistry 21, 4671-4679 (1982). [CrossRef] [PubMed]
- L. Davenport, J. R. Knutson, and L. Brand, "Excited-state proton transfer of equilenin and dihydroequilenin: interaction with bilayer vesicles," Biochemistry 25, 1186-1195 (1986). [CrossRef] [PubMed]
- I. Gautier, M. Tramier, C. Durieux, J. Coppey, R. B. Pansu, J. C. Nicolas, K. Kemnitz, and M. Coppey-Moisan, "Homo-FRET microscopy in living cells to measure monomer-dimer transition of GFP-tagged proteins," Biophys. J. 80, 3000-3008 (2001). [CrossRef] [PubMed]
- K. Kemnitz, L. Pfeifer, R. Paul, and M. Coppey-Moisan, "Novel detectors for fluorescence lifetime imaging on the picosecond time scale," J. Fluoresc. 7, 93-98. (1997). [CrossRef]
- K. Kemnitz, L. Pfeifer, R. Paul, A. Fink, and A. Bergmann, "Time- and space correlated single photon counting Spectroscopy," in Optical and Imaging Techniques for Biomonitoring, H.-J. Foth, R. Marchesini, H. Podbielska M.D., M. Robert-Nicoud, H. Schneckenburger, eds., SPIE Proc. 2628, 2-11 (1995).
- J. Lippincott-Schwartz and G. H. Patterson, "Development and use of fluorescent protein markers in living cells," Science 300, 87-90 (2003). [CrossRef] [PubMed]
- P. Kapusta, R. Erdmann, U. Ortmann, and M. Wahl, "Time-resolved fluorescence anisotropy measurements made simple," J. Fluoresc. 13, 179-183 (2003). [CrossRef]
- M. Tramier, K. Kemnitz, C. Durieux, and M. Coppey-Moisan, "Picosecond time-resolved microspectrofluorometry in live cells exemplified by complex fluorescence dynamics of popular probes ethidium and cyan fluorescent protein," J. Microsc. 213, 110-118 (2004). [CrossRef] [PubMed]
- R. R. Duncan, A. Bergmann, M. A. Cousin, D. K. Apps, and M. J. Shipston, "Multi-dimensional time-correlated single photon counting (TCSPC) fluorescence lifetime imaging microscopy (FLIM) to detect FRET in cells," J. Microsc. 215, 1-12 (2004). [CrossRef] [PubMed]
- W. Holzer, A. Penzkofer, M. Fuhrmann, and P. Hegemann, "Spectroscopic characterization of flavin mononucleotide bound to the LOV1 domain of Phot1 from Chlamydomonas reinhardtii," Photochem. Photobiol. 75, 479-487 (2002). [CrossRef] [PubMed]
- M. H. Seifert, D. Ksiazek, M. K. Azim, P. Smialowski, N. Budisa, and T. A. Holak, "Slow exchange in the chromophore of a green fluorescent protein variant," J. Am. Chem. Soc. 124, 7932-7942 (2002). [CrossRef] [PubMed]
- S. Habuchi, M. Cotlet, J. Hofkens, G. Dirix, J. Michiels, J. Vanderleyden, V. Subramaniam, and F. C. D. Schryver, "Resonance Energy Transfer in a Calcium Concentration-Dependent Cameleon Protein," Biophys. J. 83, 3499-3506 (2002). [CrossRef] [PubMed]
Cited By |
OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.





OSA is a member of 