Enhancing two-color absorption, self-phase modulation and raman microscopy signatures in tissue with femtosecond laser pulse shaping
Nonlinear microscopies (most commonly, two-photon fluorescence, second harmonic generation, and coherent anti-Stokes Raman scattering (CARS)) have had notable successes in imaging a variety of endogenous and exogenous targets in recent years. These methods generate light at a color different from any of the exciting laser pulses, which makes the signal relatively easy to detect. Our work has focused on developing microscopy techniques using a wider range of nonlinear signatures (two-photon absorption of nonfluorescent species, self phase modulation) which have some specific advantages - for example, in recent papers we have shown that we can differentiate between different types of melanin in pigmented lesions, image hemoglobin and its oxygenation, and measure neuronal activity. In general, these signatures do not generate light at a different color and we rely on the advantages of femtosecond laser pulse shaping methods to amplify the signals and make them visible (for example, using heterodyne detection of the induced signal with one of the co-propagating laser pulses). Here we extend this work to stimulated Raman and CARS geometries. In the simplest experiments, both colors arise from filtering a single fs laser pulse, then modulating afterwards; in other cases, we demonstrate that spectral reshaping can retain high frequency resolution in Raman and CARS geometries with femtosecond laser pulses. © 2009 SPIE.
