Abstract

We report on third-harmonic generation (THG) of biomolecular solutions at the fluid/glass interface as a means to probe resonant contributions to their nonlinear absorption spectra that could serve as contrast mechanisms for functional imaging. Our source was 100fs laser pulses whose center wavelength varied from 760 to 1000nm. We find evidence of a two-photon resonance in the ratio of third-order susceptibilities, χsample(3)(3ω)χglass(3), for aqueous solutions of Rhodamine B, Fura-2, and hemoglobin and a three-photon resonance in χsample(3)(3ω)χglass(3) for solutions of bovine serum albumin. Consistent with past work, we find evidence of a one-photon resonance of χsample(3)(3ω)χglass(3) for water, while confirming a lack of resonant enhancement for benzene. At physiological concentrations, hemoglobin in different ligand-binding states could be distinguished on the basis of features of its THG spectrum.

© 2006 Optical Society of America

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