High-throughput label-free nonlinear optical microscopy via frequency-gated amplitude and lifetime demodulation

Label-free nonlinear microscopy techniques, including Auto-Fluorescence (AF), Harmonic Generation (HG), and Fluorescence Lifetime Imaging Microscopy (FLIM), provide powerful tools for analyzing biological tissues. The simultaneous label-free autofluorescence multi-harmonic (SLAM) microscopy method integrates nonlinear optical contrast mechanisms to reveal molecular and structural signatures. These modalities detect intrinsic optical signals generated by ultrafast laser pulses interacting with the sample. In this presentation, we introduce a method that enables high-speed, high-SNR imaging of weak signals. Additionally, we demonstrate its capability to facilitate FLIM imaging, further enhancing its potential for biomedical research and diagnostics.