Coherent Raman Scattering (CRS) microscopy is capable of non-invasive, label-free imaging of tissues and cells, based on their intrinsic vibrational response. Coherent Anti-Stokes Raman Scattering (CARS) and Stimulated Raman Scattering (SRS) are the two most commonly employed CRS techniques. With respect to CARS, SRS presents several advantages: the signal is proportional to species concentration and is free of non-resonant four-wave-mixing. However, SRS is technically very challenging, as it requires high-speed modulation and synchronous lock-in detection to measure the tiny (10−4 or less) differential transmission (ΔT/T) signal sitting on top of a large linear background. SRS imaging has been demonstrated up to video-rate speed but at a single vibrational frequency, which provides a too limited information to distinguish different species within complex heterogeneous systems with spectrally overlapping chemical components. While broadband CARS has been reported, recording broadband SRS spectra is very challenging: so far, multiplex SRS has only been performed with narrowband pulses, by rapidly scanning the pump-Stokes frequency detuning.

© 2015 IEEE

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