High-Speed Imaging with rapidFLIM

PicoQuant GmbH

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Fluorescence Lifetime Imaging (FLIM) is a versatile microscopy method, which combines fluorescence lifetime information with spatial localization, enabling the investigation of biochemical and physical processes, detecting changes in the local environment of the sample, molecular interactions, or conformational changes via Förster Resonance Energy Transfer (FRET). FLIM data acquisition based on Time-Correlated Single Photon Counting (TCSPC) is considered a somewhat slow process, due to the time required to collect a sufficient number of photons per pixel for reliable data analysis.

The rapidFLIM approach exploits recent hardware developments such as TCSPC electronics and hybrid photomultiplier detectors with ultra short dead times. These improved hardware components enable using high detection count rates, making it possible to achieve better photon statistics in much shorter time spans. With rapidFLIM, imaging can now be performed at a rate of several frames per second.

Depending on image size and sample brightness, more than 15 images can be acquired per second, making it possible to follow dynamic processes such as transient protein interaction in living cells or chemical reactions as well as highly mobile species without sacrificing the lifetime contrast or the high spatial resolution of the confocal microscope.

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