22 February 2013 Fluorescence lifetime imaging with near-infrared dyes
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Near-infrared (NIR) dyes are used as fluorescence markers in small-animal imaging and in diffuse optical tomography of the human brain. In these applications it is important to know whether the dyes bind to proteins or other tissue constituents, and whether their fluorescence lifetimes depend on the targets they are bound to. Unfortunately, neither the lasers nor the detectors of commonly used confocal and multiphoton laser scanning microscopes allow for excitation and detection of NIR fluorescence. We therefore upgraded existing confocal TCSPC FLIM systems with NIR lasers and NIR sensitive detectors. In multiphoton systems we used the Ti:Sa laser as a one-photon excitation source in combination with an NIR-sensitive detector in the confocal beam path. We tested a number of NIR dyes in biological tissue. Some of them showed clear lifetime changes depending on the tissue structures they are bound to. We therefore believe that NIR FLIM can deliver supplementary information on the tissue constitution and on local biochemical parameters.
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Wolfgang Becker, Wolfgang Becker, Vladislav Shcheslavskiy, Vladislav Shcheslavskiy, "Fluorescence lifetime imaging with near-infrared dyes", Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 85880R (22 February 2013); doi: 10.1117/12.2003608; https://doi.org/10.1117/12.2003608

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