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For high-signal-rate nonlinear microscopy using an analog PMT to detect multiple photons per excitation pulse, the conversion of analog output to photon counts is required to realize quantitative imaging. To enable this ability, we propose a method for photon counting with analog photodetection based on “imaging” of standard dye solutions and a mathematical model of Poisson photon statistics. Using this method, we not only enable the quantification across different types of PMTs from different setups, but also quantify the channel leakage for multichannel or multispectral imaging systems. This method paves the way for analog PMTs in high-signal-rate quantitative imaging of either label-free or labeled samples.
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Geng Wang, Janet E. Sorrells, Rishyashring R. Iyer, Stephen A. Boppart, Haohua Tu, "Photon counting with analog photodetection for high-signal-rate quantitative nonlinear microscopy," Proc. SPIE PC12370, Design and Quality for Biomedical Technologies XVI, PC1237006 (6 March 2023); https://doi.org/10.1117/12.2648438