Raman microscopy based on doubly-resonant four-wave mixing (DR-FWM)

Doubly-resonant four-wave mixing (DR-FWM) is a nondegenerate four-wave mixing process in which four photons interact to coherently probe two distinct Raman resonances. We demonstrate DR-FWM microscopy as a label-free and nondestructive molecular imaging modality with high chemical specificity on the submicron scale by imaging alkyne-substituted oleic acid in both aqueous and lipid-rich environments. DR-FWM microscopy is contrasted to coherent anti-Stokes Raman scattering (CARS) microscopy and it is shown that the coherent addition of two simultaneously probed Raman resonances leads to a significant increase in signal without increasing the non-resonant background. Thus, this scheme enables the detection of weak Raman signals through amplification by a strong Raman resonance, potentially increasing the overall detection sensitivity beyond what has been demonstrated by either CARS or stimulated Raman scattering (SRS).