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Combined Fluorescence and Phosphorescence Lifetime Imaging (FLIM / PLIM) with the Zeiss LSM 710 NLO Microscopes

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The PLIM technique described here is based on modulating the titanium-sapphire laser of a multiphoton microscope by a signal synchronous with the pixel clock of the scanner, and recording the fluorescence and phosphorescence signals by multi-dimensional TCSPC [1]. Fluorescence is recorded during the onphase of the laser, phosphorescence during the off-phase. Laser modulation is achieved by controlling the the AOM of the laser scanning microscope by a signal generated in the TCSPC system.
This document contains instructions of how to set the parameters of the bh SPC-150 or SPC-830 FLIM systems for combined fluorescence (FLIM) and phosphorescence lifetime imaging (PLIM). It should be considered a supplement to the bh TCSPC Handbook [1] and the bh Handbook of the FLIM systems for the Zeiss LSM 510 and 710 laser scanning microscopes [2].

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