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September 5, 2016

rapidFLIM – redefining the standards for dynamic FLIM imaging

PicoQuant releases application note describing a novel fluorescence lifetime imaging method with fast image acquisition

A recent publication by PicoQuant lays out the basics of rapidFLIM, a novel, dynamic Fluorescence Lifetime Imaging (FLIM) method, along with practical applications. The rapidFLIM approach allows image acquisition with several frames per second and thus opens the way for the study of dynamic processes in living cells or other materials through fast fluorescence lifetime imaging. It is ideally suited for observing fast processes, like protein interaction or chemical interactions, following highly mobile species including live cells, moving organelles, or nanoparticles, as well as investigate Förster Resonance Energy Transfer (FRET) dynamics. Depending on sample brightness and image size, rapidFLIM allows acquiring more than 10 to 15 frames per second.

Time Series of diffusing beads in water - time seriesThe method exploits recent hardware developments such as TCSPC modules with ultra short dead times and appropriate hybrid photomultiplier detector assemblies enabling significantly higher detection count rates. These improved hardware components make it possible to achieve much better photon statistics in significantly shorter time spans while being able to perform FLIM imaging for fast processes in a precise manner along with high optical resolution. By using a pattern matching analysis integrated into PicoQuants SymPhoTime 64 software, excellent FLIM contrast can be obtained without need for classic decay data fitting.

The rapidFLIM method can be integrated into many existing confocal laser scanning microscopes from major manufacturers such as Nikon, Olympus, or Zeiss through LSM upgrade kits offered by PicoQuant. The required kits include a TimeHarp 260 NANO with negligible dead time and a matched hybrid detector from the PMA Hybrid Series, along with a choice of pulsed laser sources and the system software SymPhoTime 64.

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