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

Serge Blaise Pelet; Micheal John Robert Previte; Ki Hean Kim; Peter T. C. So

doi:10.1117/12.537620

21 June 2004 Development of an improved FRET-imaging two-photon microscope combining lifetime and spectral imaging

Serge Blaise Pelet, Micheal John Robert Previte, Ki Hean Kim, Peter T. C. So

Proceedings Volume 5323, Multiphoton Microscopy in the Biomedical Sciences IV; (2004) https://doi.org/10.1117/12.537620
Event: Biomedical Optics 2004, 2004, San Jose, CA, United States

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.

Citation Download Citation

Serge Blaise Pelet, Micheal John Robert Previte, Ki Hean Kim, and Peter T. C. So "Development of an improved FRET-imaging two-photon microscope combining lifetime and spectral imaging", Proc. SPIE 5323, Multiphoton Microscopy in the Biomedical Sciences IV, (21 June 2004); https://doi.org/10.1117/12.537620

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