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  3. Fast Wide-field Light Sheet Electro-optic Flim.
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  3. Fast Wide-field Light Sheet Electro-optic Flim.

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Fast wide-field light sheet electro-optic FLIM.

V Rose Knight, Nils Bode, Dara P Dowlatshahi

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    View abstract on PubMed

    Summary
    This summary is machine-generated.

    We developed a new method for volumetric fluorescence lifetime microscopy (FLIM) using electro-optic gating. This light sheet electro-optic FLIM (LS-EO-FLIM) technique enables rapid 3D imaging of biological samples.

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    Area of Science:

    • Biophotonics
    • Microscopy
    • Molecular Imaging

    Background:

    • Fluorescence Lifetime Microscopy (FLIM) provides functional and molecular information.
    • Traditional FLIM methods can be slow for volumetric imaging.
    • Selective Plane Illumination Microscopy (SPIM) offers optical sectioning for 3D imaging.

    Purpose of the Study:

    • To demonstrate volumetric FLIM using electro-optic gating in a SPIM setup.
    • To enable faster 3D FLIM acquisition.
    • To image biological samples with high spatial and temporal resolution.

    Main Methods:

    • Implemented electro-optic FLIM (EO-FLIM) with a Pockels cell.
    • Integrated EO-FLIM gating with a selective plane illumination microscope (SPIM).
    • Achieved light sheet electro-optic FLIM (LS-EO-FLIM) with an 80 MHz gating frequency and 800 µm field of view.

    Main Results:

    • Demonstrated successful volumetric imaging of fluorescent bead mixtures.
    • Acquired 3D FLIM data from live Arabidopsis thaliana root samples.
    • Utilized both genetically encoded fluorescent proteins and endogenous autofluorescence for imaging.

    Conclusions:

    • LS-EO-FLIM is a viable technique for rapid volumetric fluorescence lifetime imaging.
    • The method allows for imaging of diverse biological samples.
    • This technique advances 3D functional imaging in live biological systems.