Abstract
Abstract Lattice light-sheet microscopy (LLSM) was developed for long-term live-cell imaging with ultra-fine three-dimensional (3D) spatial resolution, high temporal resolution, and low photo-toxicity by illuminating the sample with a thin lattice-like light-sheet. Currently available schemes for generating thin lattice light-sheets often require complex optical designs. Meanwhile, limited by the bulky objective lens and optical components, the light throughput of existing LLSM systems is rather low. To circumvent the above problems, we utilize a dielectric metasurface of a single footprint to replace the conventional illumination modules used in the conventional LLSM and generate a lattice light-sheet with a ~3-fold broader illumination area and a significantly leveraged illumination efficiency, which consequently leads to a larger field of view with a higher temporal resolution at no extra cost of the spatial resolution. We demonstrate that the metasurface can manipulate spatial frequencies of an input laser beam in orthogonal directions independently to break the trade-off between the field of view and illumination efficiency of the lattice light-sheet. Compared to the conventional LLSM, our metasurface module serving as an ultra-compact illumination component for LLSM at an ease will potentially enable a finer spatial resolution with a larger numerical-aperture detection objective lens.
Highlights
Light-sheet fluorescence microscopy (LSFM) has emerged as a powerful technique for ultra-fast 3D imaging acquisitions of live samples with high spatial and temporal resolutions, low photo-toxicity and weak background noise [1,2,3,4,5,6]
Lattice light-sheet microscopy (LLSM) was developed for long-term live-cell imaging with ultra-fine three-dimensional (3D) spatial resolution, high temporal resolution, and low photo-toxicity by illuminating the sample with a thin lattice-like light-sheet
To circumvent the above problems, we utilize a dielectric metasurface of a single footprint to replace the conventional illumination modules used in the conventional LLSM and generate a lattice light-sheet with a ~3-fold broader illumination area and a significantly leveraged illumination efficiency, which leads to a larger field of view with a higher temporal resolution at no extra cost of the spatial resolution
Summary
Light-sheet fluorescence microscopy (LSFM) has emerged as a powerful technique for ultra-fast 3D imaging acquisitions of live samples with high spatial and temporal resolutions, low photo-toxicity and weak background noise [1,2,3,4,5,6]. An identical time-averaged lattice light-sheet can be obtained by scanning a line-focused laser beam over the same pupil function as that in Eric Betzig’s LLSM. In the field synthesis LLSM, the narrow light lines are generated by step-scanning a line-shaped laser beam across the back pupil plane of an objective lens. Unlike the conventional objective lens, the metasurface has the freedom of independent control of spatial frequencies in orthogonal directions, to break the limitation of illumination efficiency due to the trade-off between the thickness of light lines at the pupil and the spatial span of the resulted lattice light-sheet. Extremely narrow light lines at the pupil of the objective lens still lead to certain spans of the spatial frequency along the light-sheet dithering direction for the reason of line extension. Our demonstrations will promote to construct a highly integrated, ultracompact, mechanically stable, and user-friendly LLSM with an elaborate illumination scheme based on a dielectric metasurface
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