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Proceedings Paper

Development of an improved FRET-imaging two-photon microscope combining lifetime and spectral imaging
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Paper Abstract

FRET imaging is widely used in biology to map protein interactions in living cells. A number of recent technical developments promise to provide more accurate and quantitative measurement FRET efficiencies and protein bonding ratio. Using a reference FRET construct based on a doubly labeled DNA, one can estimate the accuracy of different techniques like spectral or lifetime imaging to resolve FRET. Further improvement have lead to the possibility of measuring in a single detector both spectral and lifetime information from the fluorescence emitted by the sample. We are currently building such a system based on an intensity modulated multi-anode PMT and have developed a global fitting algorithm to extract valuable information from combined spectral and lifetime imaging data sets.

Paper Details

Date Published: 21 June 2004
PDF: 6 pages
Proc. SPIE 5323, Multiphoton Microscopy in the Biomedical Sciences IV, (21 June 2004); doi: 10.1117/12.537620
Show Author Affiliations
Serge Blaise Pelet, Massachusetts Institute of Technology (United States)
Micheal John Robert Previte, Massachusetts Institute of Technology (United States)
Ki Hean Kim, Massachusetts Institute of Technology (United States)
Peter T. C. So, Massachusetts Institute of Technology (United States)


Published in SPIE Proceedings Vol. 5323:
Multiphoton Microscopy in the Biomedical Sciences IV
Ammasi Periasamy; Peter T. C. So, Editor(s)

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