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Journal of Biomedical Optics • Open Access

Innovating lifetime microscopy: a compact and simple tool for life sciences, screening, and diagnostics
Author(s): Alessandro Esposito; Hans Gerritsen; Thierry Oggier; Felix Lustenberger; Fred S. Wouters

Paper Abstract

Fluorescence lifetime imaging microscopy (FLIM) allows the investigation of the physicochemical environment of fluorochromes and protein-protein interaction mapping by Förster resonance energy transfer (FRET) in living cells. However, simpler and cheaper solutions are required before this powerful analytical technique finds a broader application in the life sciences. Wide-field frequency-domain FLIM represents a solution whose application is currently limited by the need for multichannel-plate image intensifiers. We recently showed the feasibility of using a charge-coupled device/complementory metal-oxide semiconductor (CCD/CMOS) hybrid lock-in imager, originally developed for 3-D vision, as an add-on device for lifetime measurements on existing wide-field microscopes. In the present work, the performance of the setup is validated by comparison with well-established wide-field frequency-domain FLIM measurements. Furthermore, we combine the lock-in imager with solid-state light sources. This results in a simple, inexpensive, and compact FLIM system, operating at a video rate and capable of single-shot acquisition by virtue of the unique parallel retrieval of two phase-dependent images. This novel FLIM setup is used for cellular and FRET imaging, and for high-throughput and fast imaging applications. The all-solid-state design bridges the technological gap that limits the use of FLIM in areas such as drug discovery and medical diagnostics.

Paper Details

Date Published: 1 May 2006
PDF: 8 pages
J. Biomed. Opt. 11(3) 034016 doi: 10.1117/1.2208999
Published in: Journal of Biomedical Optics Volume 11, Issue 3
Show Author Affiliations
Alessandro Esposito, GWDG (Germany)
Hans Gerritsen, Hortus Group (United States)
Thierry Oggier, Ctr. Suisse d'Electronique et de Microtechnique SA (Switzerland)
Felix Lustenberger, Ctr. Suisse d'Electronique et de Microtechnique SA (Switzerland)
Fred S. Wouters, European Neuroscience Institute (Germany)

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