Coherent anti-Stokes Raman scattering (CARS) microscopy is a powerful technique to image the chemical composition
of complex samples in biophysics, biology and materials science. CARS is a four-wave mixing process. The application
of a spectrally narrow pump beam and a spectrally wide Stokes beam excites multiple Raman transitions, which are
probed by a probe beam. This generates a coherent directional CARS signal with several orders of magnitude higher
intensity relative to spontaneous Raman scattering. Recent advances in the development of ultrafast lasers, as well as
photonic crystal fibers (PCF), enable multiplex CARS. In this study, we employed two scanning imaging methods. In
one, the detection is performed by a photo-multiplier tube (PMT) attached to the spectrometer. The acquisition of a
series of images, while tuning the wavelengths between images, allows for subsequent reconstruction of spectra at each
image point. The second method detects CARS spectrum in each point by a cooled coupled charged detector (CCD)
camera. Coupled with point-by-point scanning, it allows for a hyperspectral microscopic imaging. We applied this
CARS imaging system to study biological samples such as oocytes.
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