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Coherent anti-Stokes Raman scattering (CARS) spectra have been obtained for ferrocytochrome c and cyano cobalamin in aqueous solution at millimolar concentrations, using a pair of tunable dye lasers pumped by a pulsed nitrogen laser. Resonance enhancement was obtained by tuning the omega1 laser to the visible absorption bands of the samples. The spectral features correspond to those observed in the conventional resonance Raman spectra. It appears that CARS spectroscopy, with its advantageous fluorescence rejection, can be usefully applied to biological samples by exploiting resonance enhancement. While the background scattering from water is 10 times higher than that of benzene and other aromatic solvents, it is actually at the low end of the scale for most liquids. The anomalously low background of aromatic liquids is thought to result from competition by the unusually efficient stimulated Raman scattering which they display. Off-resonance spectra for both cobalamin and cytochrome c contain negative peaks, i.e., absorption bands in the background. These are interpreted as inverse Raman processes induced by the omega1 photons in the presence of the continuum provided by the background scattering. While both CARS and the inverse Raman effect are subject to resonance enhancement, the wavelength dependence of CARS is evidently steeper.

Coherent anti-Stokes Raman scattering (CARS) spectra, with resonance enhancement, of cytochrome c and vitamin B12 in dilute aqueous solution.