Research Papers

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

[+] Author Affiliations
Alessandro Esposito

European Neuroscience Institute, Göttingen, Cell Biophysics Group, Waldweg 33, D-37073 Göttingen, Germany and Utrecht University, Debye Institute, P. O. Box 80.000, NL 3508 TA, Utrecht, The Netherlands

Hans C. Gerritsen

Utrecht University, Debye Institute, P. O. Box 80.000, NL 3508 TA, Utrecht, The Netherlands

Thierry Oggier, Felix Lustenberger

CSEM SA, Image Sensing Group, Photonics Division, Badenerstrasse 569, CH-8048 Zurich, Switzerland

Fred S. Wouters

European Neuroscience Institute, Göttingen, Cell Biophysics Group, Waldweg 33, D-37073 Göttingen, Germany

J. Biomed. Opt. 11(3), 034016 (June 07, 2006). doi:10.1117/1.2208999
History: Received November 16, 2005; Revised March 02, 2006; Accepted March 03, 2006; Published June 07, 2006
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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.

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© 2006 Society of Photo-Optical Instrumentation Engineers

Citation

Alessandro Esposito ; Hans C. Gerritsen ; Thierry Oggier ; Felix Lustenberger and Fred S. Wouters
"Innovating lifetime microscopy: a compact and simple tool for life sciences, screening, and diagnostics", J. Biomed. Opt. 11(3), 034016 (June 07, 2006). ; http://dx.doi.org/10.1117/1.2208999


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