Abstract
We present a novel time-correlated single photon counting (TCPSC) imaging technique that allows time-resolved multi-wavelength imaging in conjunction with a laser scanning microscope and a pulsed excitation source. The technique is based on a four-dimensional histogramming process that records the photon density over time, the x-y coordinates of the scanning area and the detector channel number. The histogramming process avoids any time gating or wavelength scanning and therefore yields a near-perfect counting efficiency. Applied to resonance energy transfer (RET) measurements, the setup is capable to record time-resolved fluorescence decays for the donor and the acceptor simultaneously.
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