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25 February 2009High performance multimodal CARS microscopy using a single femtosecond source
We demonstrate high performance coherent anti-Stokes Raman scattering (CARS) microscopy using a single
femtosecond Ti:Sapphire laser source combined with a photonic crystal fiber (PCF). By adjusting the chirp of
the femtosecond pump and Stokes laser pulses, we achieve high quality multimodal imaging (simultaneous CARS,
two-photon fluorescence, and second harmonic generation) of live cells and tissues. The tuneable Ti:sapphire
output provides the pump beam directly, while part of this is converted to the red-shifted Stokes pulse using a
PCF having two close-lying zero dispersion wavelengths. This PCF gives good power and stability over Stokes
shifts ranging from below 2300 cm-1 to over 4000 cm-1. This tuning range can be accessed by simply controlling
the time delay between the input pulses. This allows fast, continuous computer-controlled tuning of the Stokes
shift over a broad range, without involving any adjustment of either the femtosecond laser or the PCF. The
simultaneous optimization of CARS, two-photon fluorescence and second harmonic generation is achieved by
controlling the degree of chirp and involves a trade-off between spectral resolution of the CARS process and
signal strength. This is illustrated by showing applications of the multimodal CARS imaging and optimization
technique to biomedical problems involving both live cells and tissues.
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Adrian F. Pegoraro, Andrew Ridsdale, Douglas J. Moffatt, John P. Pezacki, Albert Stolow, "High performance multimodal CARS microscopy using a single femtosecond source," Proc. SPIE 7183, Multiphoton Microscopy in the Biomedical Sciences IX, 71830Z (25 February 2009); https://doi.org/10.1117/12.809560