Compact plane illumination plugin device to enable light sheet fluorescence imaging of multi-cellular organisms on an inverted wide-field microscope

Abstract

Abstract:We developed a compact plane illumination plugin (PIP) device which enabled plane illumination and light sheet fluorescence imaging on a conventional inverted microscope. The PIP device allowed the integration of microscope with tunable laser sheet profile, fast image acquisition, and 3-D scanning. The device is both compact, measuring approximately 15 by 5 by 5 cm, and cost-effective, since we employed consumer electronics and an inexpensive device molding method. We demonstrated that PIP provided significant contrast and resolution enhancement to conventional microscopy through imaging different multi-cellular fluorescent structures, including 3-D branched cells in vitro and live zebrafish embryos. Imaging with the integration of PIP greatly reduced out-of-focus contamination and generated sharper contrast in acquired 2-D plane images when compared with the stand-alone inverted microscope. As a result, the dynamic fluid domain of the beating zebrafish heart was clearly segmented and the functional monitoring of the heart was achieved. Furthermore, the enhanced axial resolution established by thin plane illumination of PIP enabled the 3-D reconstruction of the branched cellular structures, which leads to the improvement on the functionality of the wide field microscopy. 1938-1940 (2007

Integrating light-sheet imaging with virtual reality to recapitulate developmental cardiac mechanics

“…In step 1, we applied in-house LSFM systems (26,27,29,31) to acquire image stacks of cardiac events. For the large-scale samples, such as the adult mouse hearts and hydrogel scaffolds, dual-sided laser beams were applied to evenly illuminate the samples, followed by orthogonal fluorescent or autofluorescent detection (Sup- Scale: 1-5 (1, strongly disagree; 5, strongly agree).…”

“…The details of the in-house LSFM systems have been covered by existing publications (26,27,29,31). Our light-sheet imaging systems enable us to increase the working distance with sufficient spatial resolution needed to track the organ development.…”

“…Unlike VR scenarios that are commonly generated by computer graphics or a 360-degree camera (11,12), the advent of light-sheet fluorescence microscopy (LSFM) (13)(14)(15)(16)(17)(18)(19)(20)(21) allows for capturing of physiological events in the 3D or 4D domain that can be further adapted to VR headsets, such as Google Cardboard or Daydream, for effective VR visualization. In comparison with wide-field or confocal microscopy, LSFM enables multidimensional imaging at the single-cell resolution (22)(23)(24)(25)(26)(27)(28)(29), which is critical for integrating high-fidelity physiological research with VR visualization. 4D LSFM further enables in vivo imaging of contracting embryonic hearts in zebrafish (Danio rerio) (30,31), Caenorhabditis elegans (32,33), and Drosophila melanogaster (34,35).…”

Three-dimensional, isotropic imaging of mouse brain using multi-view deconvolution light sheet microscopy

“…[1][2][3] LSFM is fundamentally characterized by setting a separated plane illumination orthogonal to the wide-¯eld detection, thus enabling optical sectioning inside the samples with sharp contrast and high axial resolution superior to conventional microscopes. [4][5][6][7] In the past decade, besides the basic Gaussian light-sheet generated with a cylindrical lens, LSFM have derived a series of structural variants, including multidirectional SPIM (mSPIM), Digital Scanned Laser Light-sheet Fluorescence Microscopy (DSLM), Bessel beam plane illumination microscopy and so on. [8][9][10][11][12][13][14] In accordance to these advanced LSFM imaging modalities, a variety of novel biomedical studies have been accomplished based on their excellent visualization.…”

MEMS micromirror based light sheet generator for biomedical imaging

“…Two general concepts are commonly used. The creation of a light sheet through the use of cylindrical lenses, with a compact concept being demonstrated recently [2], or the use of scanned single point laser beams [3]. The advantages of using scanning systems are the extra flexibility for tailored dimensions of the light sheet and the potential for easier synchronization control.…”