Two-color Stimulated Raman scattering (SRS) microscopy has shown great potential in label-free digital histology with diagnostic results similar to H&E stain. However, achieving real-time two-color SRS imaging is challenging. We have precisely engineered the pulse profiles of the Stokes beams, and fully utilized the in-phase (X) and quadrature (Y) outputs of a phase sensitive lock-in amplifier to realize simultaneous two-color SRS imaging. We have demonstrated its robustness and advantages in rapid histology, as well as real-time in vivo imaging of live animals, both in transmission and epi modes. Moreover, we have also adapted this method to other pump-probe based microscopes, such as transient absorption (TA) microscopy.
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