Leica have recently introduced hybrid detectors for the non-descanned (RLD) ports of their SP5 and SP8 multiphoton laser scanning microscopes. We have tested these detectors for FLIM with the bh TCSPC modules. We describe the TCSPC parameter setup and operating conditions for the detectors, and demonstrate the performance for typical samples. For more information please see application note ‘Multiphoton FLIM with the Leica HyD RLD Detectors‘.
Left: Lifetime image of a BPAE cell, labelled with Alexa 488 and Mito Tracker red. Amplitude-weighted lifetime of double-exponential decay. Right: Fluorescence decay curve at position marked with red cross. bh SPC-830 TCSPC module, image format 512 x 512 pixels, 256 time channels. Analysis by bh SPCImage, double exponential decay model with incomplete decay option.