A versatile tiling light sheet microscope for cleared tissues imaging

AbstractWe present a tiling light sheet microscope compatible with all tissue clearing methods for rapid multicolor 3D imaging of centimeter-scale cleared tissues with micron-scale (4×4×10 μm3) to nanometer-scale (70×70×200 nm3) spatial resolution. The microscope uses tiling light sheets to achieve a more advanced multicolor 3D imaging ability than conventional light sheet microscopes. The illumination light is phase modulated to adjust the 3D imaging ability of the microscope based on the tissue size and the desired spatial resolution and imaging speed, and also to keep the microscope aligned. The ability of the microscope to align via phase modulation alone also makes the microscope reliable and easy to operate. Here we describe the working principle and design of the microscope. We demonstrate its imaging ability by imaging various cleared tissues prepared by different tissue clearing and tissue expansion techniques.

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Tiling light sheet selective plane illumination microscopy using discontinuous light sheets

10.1101/378232 ◽

2018 ◽

Cited By ~ 1

Author(s):

Liang Gao

Keyword(s):

Spatial Resolution ◽

3D Imaging ◽

High Spatial Resolution ◽

Image Data ◽

Image Plane ◽

Light Sheet ◽

Selective Plane Illumination Microscopy ◽

Novel Method ◽

Technical Details ◽

Imaging Speed

AbstractTiling light sheet selective plane illumination microscopy (TLS-SPIM) improves 3D imaging ability of SPIM by using a real-time optimized tiling light sheet. However, the imaging speed decreases, and size of the raw image data increases proportionally to the number of tiling positions in TLS-SPIM. The decreased imaging speed and the increased raw data size could cause significant problems when TLS-SPIM is used to image large specimens at high spatial resolution. Here, we present a novel method to solve the problem. Discontinuous light sheets created by scanning coaxial beam arrays synchronized with camera exposures are used for 3D imaging to decrease the number of tiling positions required at each image plane without sacrificing the spatial resolution. We investigate the performance of the method via numerical simulation and discuss the technical details of the method.

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In-vivo 3D imaging of Zebrafish’s intersegmental vessel development by a bi-directional light-sheet illumination microscope

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2021.03.160 ◽

2021 ◽

Vol 557 ◽

pp. 8-13

Author(s):

Xiaofei Qin ◽

Chong Chen ◽

Linbo Wang ◽

Xiaohu Chen ◽

Yong Liang ◽

...

Keyword(s):

3D Imaging ◽

Light Sheet ◽

Vessel Development

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3D spatial resolution and spectral resolution of interferometric 3D imaging spectrometry

Applied Optics ◽

10.1364/ao.55.002489 ◽

2016 ◽

Vol 55 (10) ◽

pp. 2489 ◽

Cited By ~ 6

Author(s):

Masaki Obara ◽

Kyu Yoshimori

Keyword(s):

Spatial Resolution ◽

Spectral Resolution ◽

3D Imaging ◽

Imaging Spectrometry

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High-resolution medical imaging system for 3D imaging of radioactive sources with 1-mm FWHM spatial resolution

10.1117/12.480393 ◽

2003 ◽

Cited By ~ 2

Author(s):

Robert K. Smither

Keyword(s):

High Resolution ◽

Medical Imaging ◽

Spatial Resolution ◽

3D Imaging ◽

Imaging System ◽

Radioactive Sources

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Advanced Synchrotron Radiation and Neutron Scattering Techniques for Microstructural Characterization in Industrial Research

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.750.53 ◽

2017 ◽

Vol 750 ◽

pp. 53-66

Author(s):

Fabrizio Fiori ◽

Emmanuelle Girardin ◽

Alessandra Giuliani ◽

Adrian Manescu ◽

Serena Mazzoni ◽

...

Keyword(s):

Residual Stresses ◽

Spatial Resolution ◽

Small Angle ◽

3D Imaging ◽

Volume Fraction ◽

Rapid Development ◽

Small Angle Scattering ◽

Bragg Diffraction ◽

X Rays ◽

Angle Scattering

The rapid development of new materials and their application in an extremely wide variety of research and technological fields has lead to the request of increasingly sophisticated characterization methods. In particular residual stress measurements by neutron diffraction, small angle scattering of X-rays and neutrons, as well as 3D imaging techniques with spatial resolution at the micron or even sub-micron scale, like micro-and nano-computerized tomography, have gained a great relevance in recent years.Residual stresses are autobalancing stresses existing in a free body not submitted to any external surface force. Several manufacturing processes, as well as thermal and mechanical treatments, leave residual stresses within the components. Bragg diffraction of X-rays and neutrons can be used to determine residual elastic strains (and then residual stresses by knowing the material elastic constants) in a non-destructive way. Small Angle Scattering of neutrons or X-rays, complementary to Transmission Electron Microscopy, allows the determination of structural features such as volume fraction, specific surface and size distribution of inhomogeneities embedded in a matrix, in a huge variety of materials of industrial interest. X-ray microtomography is similar to conventional Computed Tomography employed in Medicine, allowing 3D imaging of the investigated samples, but with a much higher spatial resolution, down to the sub-micron scale. Some examples of applications of the experimental techniques mentioned above are described and discussed.

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Fluorogenic probe for fast 3D whole-cell DNA-PAINT

10.1101/2020.04.29.066886 ◽

2020 ◽

Cited By ~ 3

Author(s):

Kenny KH Chung ◽

Zhao Zhang ◽

Phylicia Kidd ◽

Yongdeng Zhang ◽

Nathan D Williams ◽

...

Keyword(s):

3D Imaging ◽

Super Resolution ◽

Fold Increase ◽

High Fidelity ◽

Optical Sectioning ◽

High Background ◽

Fluorogenic Probe ◽

Microscopy Method ◽

Super Resolution Microscopy ◽

Imaging Speed

AbstractDNA-PAINT is an increasingly popular super-resolution microscopy method that can acquire high-fidelity images at nanometer resolution. It suffers, however, from high background and very slow imaging speed, both of which can be attributed to the presence of unbound fluorophores in solution. We present a fluorogenic DNA-PAINT probe that solves these problems and demonstrate 3D imaging without the need for optical sectioning and a 26-fold increase in imaging speed over regular DNA-PAINT.

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