rapidFLIM – Redefining the Standards for Dynamic FLIM Imaging
Novel Microscopy Technique for Visualizing Fast Rrocesses
Fluorescence Lifetime Imaging (FLIM) is a versatile microscopy method where fluorescence lifetime information is combined with spatial localization, allowing investigating 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 is typically based on Time-Correlated Single Photon Counting (TCSPC) electronics, pulsed diode lasers, and highly sensitive single photon counting detectors. Up to now TCSPC data acquisition is considered a somewhat slow process, due to the time required to collect a sufficient number of photons per pixel for reliable data analysis.
rapidFLIM allows acquiring several FLIM images per second by exploiting recent hardware developments such as TCSPC modules with ultra short dead times and hybrid photomultiplier detectors. These improved hardware components enable higher detection count rates, making it possible to achieve much better photon statistics in shorter time spans. Thus, FLIM imaging can be performed for dynamic processes (e.g., transient protein interaction in living cells or chemical reaction) as well as highly mobile species in a precise manner and with high optical resolution. As an additional benefit, the accuracy of the data analysis is comparable to that of conventional TCSPC.