We show the applicability of vertex component analysis (VCA) of hyperspectral CARS images in generating a similar contrast profile to that obtained in “multimodal imaging” that uses signals from three separate nonlinear optical techniques. Using an atherosclerotic rabbit aorta test image, we show that the VCA algorithm provides pseudocolor contrast that is comparable to multimodal imaging, thus suggesting that under certain conditions much of the information gleaned from a multimodal nonlinear optical approach can be sufficiently extracted from the CARS hyperspectral stack itself. This is useful for unsupervised contrast generation on hyperspectral CARS implementations such as multiplex CARS that may not have multimodal capabilities. The utility of VCA as a quantitative analysis tool in CARS is also addressed.

Abstract:We demonstrate spectral-focusing based coherent anti-Stokes Raman scattering (SF-CARS) hyper-microscopy capable of probing vibrational frequencies from 630 cm 1 to 3250 cm 1 using a single Ti:Sapphire femtosecond laser operating at 800 nm, and a commercially-available supercontinuum-generating fibre module. A broad Stokes supercontinuum with significant spectral power at wavelengths between 800 nm and 940 nm is generated by power tuning the fibre module using atypically long and/or chirped ~200 fs pump pulses, allowing convenient access to lower vibrational frequencies in the fingerprint spectral region. This work significantly reduces the instrumental and technical requirements for multimodal CARS microscopy, while expanding the spectral capabilities of an established approach to SF-CARS.

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We present a simple technique that significantly enhances the interaction of pump pulses with a supercontinuum Stokes generated by a particular nonlinear fiber for time-gated experiments such as coherent anti-Stokes Raman scattering (CARS). The enhancement is achieved through a synchronized power-tuning/time delay scheme that we call spectral surfing. In this Letter, we introduce spectral surfing and demonstrate how its application to an economical CARS hypermicroscopy scheme increases the brightness, contrast, and spectral scanning range, while potentially reducing sample light exposure.

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