Fluorescence lifetime microscopy with a time- and space-resolved single-photon counting detector

We have recently developed a wide-field photon-counting detector (the H33D detector) having high-temporal and high-spatial resolutions and capable of recording up to 500,000 photons per sec. Its temporal performance has been previously characterized using solutions of fluorescent materials with diff...

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Bibliographic Details
Published inProceedings of SPIE, the international society for optical engineering Vol. 6372
Main Authors Michalet, X, Siegmund, O H W, Vallerga, J V, Jelinsky, P, Pinaud, F F, Millaud, J E, Weiss, S
Format Journal Article
LanguageEnglish
Published United States 01.01.2006
Subjects
live cell
microchannel plate
single-molecule
fluorescence
single-photon
lifetime
quantum dot
FLIM
position sensitive anode
photocathode
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Summary:We have recently developed a wide-field photon-counting detector (the H33D detector) having high-temporal and high-spatial resolutions and capable of recording up to 500,000 photons per sec. Its temporal performance has been previously characterized using solutions of fluorescent materials with different lifetimes, and its spatial resolution using sub-diffraction objects (beads and quantum dots). Here we show its application to fluorescence lifetime imaging of live cells and compare its performance to a scanning confocal TCSPC approach. With the expected improvements in photocathode sensitivity and increase in detector throughput, this technology appears as a promising alternative to the current lifetime imaging solutions.
ISSN:0277-786X
DOI:10.1117/12.686429