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Functional in vivo imaging using fluorescence lifetime light-sheet microscopy.

Claire A Mitchell, Simon P Poland, James Seyforth

    Optics Letters
    |April 1, 2017
    PubMed
    Summary
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    We integrated fluorescence lifetime imaging microscopy (FLIM) into light-sheet microscopy for rapid, functional volumetric imaging. This novel system enables high-speed in vivo imaging with concentration-independent contrast.

    Area of Science:

    • Biophotonics
    • Microscopy
    • Zebrafish Imaging

    Background:

    • Light-sheet microscopy excels at fast, low-phototoxicity volumetric imaging.
    • Standard light-sheet microscopy primarily offers structural or concentration data.
    • Fluorescence lifetime imaging microscopy (FLIM) provides functional contrast but is often slow and complex.

    Purpose of the Study:

    • To develop a light-sheet microscopy system with integrated, high-speed FLIM capabilities.
    • To enable rapid acquisition of volumetric data with functional contrast.
    • To overcome limitations of current FLIM implementations in speed and complexity.

    Main Methods:

    • Incorporation of a dedicated frequency domain CMOS FLIM camera.
    • Integration of an intensity-modulated laser into a light-sheet setup.

    Related Experiment Videos

  • Application of the system for in vivo imaging of live transgenic zebrafish.

    • Main Results:

    • Successful addition of FLIM functionality to a light-sheet microscope.
    • Demonstration of rapid volumetric FLIM data acquisition.
    • Acquisition of concentration-independent contrast in volumetric images.

    Conclusions:

    • The developed system enables fast, functional volumetric imaging using light-sheet FLIM.
    • This approach allows for rapid in vivo FLIM data collection from biological samples.
    • The system offers a powerful tool for advanced live biological imaging.