Simultaneous two-color stimulated Raman scattering microscopy by adding a fiber amplifier to a 2 ps OPO-based SRS microscope.

Stimulated Raman scattering (SRS) microscopy is a label-free chemical imaging technique. Two-color imaging is often necessary to determine the distribution of chemical species in SRS microscopy. Current multi-color SRS imaging methods involve complicated instrumentation or longer data acquisition time or are limited to transmission imaging. In this Letter, we show that by adding a simple fiber amplifier to a 2 ps laser source and optical-parametric-oscillator-based SRS setup, one can achieve simultaneous two-color or frequency modulation SRS microscopy. The fiber amplifier can generate a wavelength tunable laser of ±10  nm around the Stokes laser wavelength at 1031 nm with average power greater than 200 mW. In vivo and ex vivo lipid-protein imaging of mouse brain and skin is demonstrated. To further demonstrate the potential of this technique in high-speed in vivo imaging, white blood cells in a blood stream are imaged in a live mouse.