Innovating lifetime microscopy: a compact and simple tool for life sciences, screening, and diagnostics

Alessandro Esposito; Hans Gerritsen; Thierry Oggier; Felix Lustenberger; Fred S. Wouters

doi:10.1117/1.2208999

1 May 2006 Innovating lifetime microscopy: a compact and simple tool for life sciences, screening, and diagnostics

Alessandro Esposito, Hans Gerritsen, Thierry Oggier, Felix Lustenberger, Fred S. Wouters

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Journal of Biomedical Optics, Vol. 11, Issue 3, 034016 (May 2006). https://doi.org/10.1117/1.2208999

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.

©(2006) Society of Photo-Optical Instrumentation Engineers (SPIE)

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Alessandro Esposito, Hans Gerritsen, Thierry Oggier, Felix Lustenberger, and Fred S. Wouters "Innovating lifetime microscopy: a compact and simple tool for life sciences, screening, and diagnostics," Journal of Biomedical Optics 11(3), 034016 (1 May 2006). https://doi.org/10.1117/1.2208999

Published: 1 May 2006

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