Abstract

Abstract Light-sheet fluorescent microscopy has become the leading technique for in vivo imaging in the fields of disease, medicine, and cell biology research. However, designing proper illumination for high image resolution and optical sectioning is challenging. Another issue is geometric constraints arising from the multiple bulky components for illumination and detection. Here, we demonstrate that those issues can be well addressed by integrating nanophotonic meta-lens as the illumination component for LSFM. The meta-lens is composed of 800-nm-thick GaN nanostructures and is designed for a light-sheet well-adapted to biological specimens such as the nematode Caenorhabditis elegans (C. elegans). With the meta-lens, the complexity of the LSFM system is significantly reduced, and it is capable of performing multicolor fluorescent imaging of live C. elegans with cellular resolution. Considering the miniature size and plane geometry of the meta-lens, our system enables a new design for LSFM to acquire in vivo images of biological specimens with high resolution.

Highlights

  • Fluorescence imaging of fine structures in live specimens provides a powerful way to understand cellular and subcellular dynamics in biology and clinical applications

  • To demonstrate the imaging capability of the lightsheet fluorescent microscopy (LSFM) system equipped with proposed light-sheet metalens, fluorescent imaging of cellular structures in live C. elegans was performed (Figure 1a)

  • A well-designed light-sheet is essential to achieve good optical sectioning as well as low out-of-focus noise. Another issue is associated with the difficulty of realizing an LSFM system capable of efficiently fixing, identifying, and tracking tiny C. elegans

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Summary

Introduction

Fluorescence imaging of fine structures in live specimens provides a powerful way to understand cellular and subcellular dynamics in biology and clinical applications. Among current microscopic imaging systems, lightsheet fluorescent microscopy (LSFM) [1–6] has become the leading technique for this purpose in recent years. Fluorescence images from the illuminated section can be observed along the detection axis, which is orthogonal to the light-sheet excitation plane. Due to the unique orthogonal scheme between excitation and collection, LSFM has multiple advantages, including a large field of view (FoV), high image resolution, and low photo-damage [3, 4, 11, 12]. This efficient imaging technique has led to numerous cutting-edge findings and helped solve many problems in various fields [4, 12–14].

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