Recent Progress in Light Sheet Microscopy for Biological Applications

Abstract

The introduction of light sheet fluorescence microscopy (LSFM) has overcome the challenges in conventional optical microscopy. Among the recent breakthroughs in fluorescence microscopy, LSFM had been proven to provide a high three-dimensional spatial resolution, high signal-to-noise ratio, fast imaging acquisition rate, and minuscule levels of phototoxic and photodamage effects. The aforementioned auspicious properties are crucial in the biomedical and clinical research fields, covering a broad range of applications: from the super-resolution imaging of intracellular dynamics in a single cell to the high spatiotemporal resolution imaging of developmental dynamics in an entirely large organism. In this review, we provided a systematic outline of the historical development of LSFM, detailed discussion on the variants and improvements of LSFM, and delineation on the most recent technological advancements of LSFM and its potential applications in single molecule/particle detection, single-molecule super-resolution imaging, imaging intracellular dynamics of a single cell, multicellular imaging: cell-cell and cell-matrix interactions, plant developmental biology, and brain imaging and developmental biology.

The Applications of Lattice Light-Sheet Microscopy for Functional Volumetric Imaging of Hippocampal Neurons in a Three-Dimensional Culture System

“…To visualize the neurons in the 3D culture system by conventional fluorescence microscopy, strong scattering from thick samples leads to blurred fluorescence signals (Movie S2 in SI). The limitation of scattering and light penetration can be improved by the use of light-sheet excitation, which offers several advantages for monitoring the neuronal activity in a 3D culture system, including a reduction in photobleaching, phototoxicity, and scattering [26,27,38]. In the original LLSM setup, two orthogonally aligned water dipping objectives were immersed in an imaging medium bath with a volume of ~10 mL [26].…”

“…Recently, selective plane illumination-based techniques have been demonstrated to be capable of high-speed volumetric imaging, [24,25]. Among them, lattice light-sheet microscopy (LLSM) has been shown to offer several advantages over other volumetric imaging tools, including less photobleaching and phototoxicity and better subcellular imaging resolution [26,27]. In LLSM, a pair of microscope objectives with perpendicular orientation shares the same focal point, where the sample is placed.…”

3D Culture Histology Cryosectioned Well Insert Technology Preserves the Structural Relationship between Cells and Biomaterials for Time‐Lapse Analysis of 3D Cultures

“…Other in vivo imaging technologies such as ultrasound, computed tomography, magnetic resonance imaging, optical coherence tomography, and light sheet microscopy are unfortunately not easily adaptable for cell culture plates. The better‐suited confocal and multiphoton microscopes provide sufficient data but they cannot be performed with histology.…”

Organoid technology in female reproductive biomedicine

“…Providing a scaffold of biomaterials along with combining microfluidics and organoid technologies may improve tissue architecture. Recent advances in CRISPR/Cas9 and gene editing technologies [149], and single-cell RNA-seq technologies [150] as well as 3D live-cell imaging using light sheet microscopy [151] will greatly accelerate applicability of organoids in biomedical research.…”