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Fluorescence time-resolved macroimaging.

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    We developed a new system for fluorescence lifetime imaging microscopy (FLIM) that can image large biological samples up to 18 mm. This technique overcomes the field-of-view limitations of traditional FLIM for in vivo biomedical research.

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    Area of Science:

    • Biomedical Imaging
    • Microscopy
    • Cell Biology

    Background:

    • Laser scanning fluorescence lifetime imaging (FLIM) is valuable for cell biology.
    • Traditional FLIM has a limited field of view (<1 mm), restricting its use for large biological samples in biomedical applications.

    Purpose of the Study:

    • To develop a FLIM system capable of imaging macroscopic samples.
    • To overcome the field-of-view limitations of conventional FLIM systems.

    Main Methods:

    • Developed a novel laser scanning FLIM system.
    • Achieved imaging of macroscopic samples up to 18 mm.
    • Maintained a lateral resolution of 15 μm.

    Main Results:

    • Successfully performed FLIM on macroscopic samples.
    • Demonstrated the system's performance using in vivo mouse tumor imaging.
    • Verified FLIM of endogenous reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and mKate2 fluorescent protein.

    Conclusions:

    • The developed FLIM system enables imaging of large biological samples.
    • This advancement expands the utility of FLIM for in vivo biomedical research.
    • The system provides high lateral resolution for macroscopic FLIM.