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Fast-Acquisition Multiphoton FLIM with the Zeiss LSM 880 NLO

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This application note demonstrates the performance of the bh FASTAC (fast acquisition) FLIM system in combination with the Zeiss LSM 880 NLO multiphoton laser scanning microscopes. The system is based on fast distribution of the photon pulses of a PMH-100 hybrid detector into four parallel TCSPC FLIM channels. The principle strongly reduces counting loss and pile up effects. If the sample allows, images can be recorded in excess of 10 MHz count rate, and at acquisition times down to the minimum frame times of the Zeiss LSM 880. Importantly, the system makes no compromises in terms of time resolution, time channel width, time channel number, or pixel number. The IRF width with fast detectors is less than 25 ps FWHM, and the temporal data are recorded with typically 1024 time channels per pixel. The time channel width can be made as small as 0.8 ps.

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