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Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy
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Simultaneous two-color stimulated Raman scattering microscopy by adding a fiber amplifier to a 2 ps OPO-based SRS

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    This study introduces a simpler method for two-color stimulated Raman scattering (SRS) microscopy using a fiber amplifier. This technique enables faster, label-free chemical imaging of biological samples, including in vivo applications.

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    Area of Science:

    • Biophotonics
    • Chemical Imaging
    • Microscopy

    Background:

    • Stimulated Raman scattering (SRS) microscopy offers label-free chemical imaging.
    • Multi-color SRS imaging is crucial for chemical species distribution but often complex.
    • Existing methods face challenges with instrumentation, acquisition time, or imaging mode.

    Purpose of the Study:

    • To develop a simplified simultaneous two-color or frequency modulation SRS microscopy technique.
    • To enhance SRS imaging capabilities by reducing complexity and acquisition time.
    • To demonstrate the applicability of the improved technique for in vivo and ex vivo imaging.

    Main Methods:

    • Integration of a fiber amplifier with a picosecond laser source and optical parametric oscillator-based SRS setup.
    • Generation of a wavelength-tunable laser (±10 nm around 1031 nm) with >200 mW average power.
    • Implementation of simultaneous two-color and frequency modulation SRS microscopy.

    Main Results:

    • Successful demonstration of simultaneous two-color SRS microscopy.
    • Achieved label-free lipid-protein imaging in mouse brain and skin (in vivo and ex vivo).
    • High-speed in vivo imaging of white blood cells in a live mouse bloodstream.

    Conclusions:

    • The enhanced SRS setup provides a simpler, faster approach for multi-color chemical imaging.
    • The technique is suitable for diverse biological applications, including dynamic in vivo studies.
    • This advancement broadens the utility of SRS microscopy for biological research.