scholarly journals Scalable volumetric imaging for ultrahigh-speed brain mapping at synaptic resolution

2017 ◽  
Author(s):  
Hao Wang ◽  
Qingyuan Zhu ◽  
Lufeng Ding ◽  
Yan Shen ◽  
Chao-Yu Yang ◽  
...  
Keyword(s):  
Brain Mapping ◽  
High Speed ◽  
Large Scale ◽  
Light Sheet ◽  
Volumetric Imaging ◽  
Tissue Sections ◽  
Light Sheet Microscopy ◽  
Cell Type Specific ◽  
Microscopy Method ◽  
Immunofluorescence Labeling

We describe a new light-sheet microscopy method for fast, large-scale volumetric imaging. Combining synchronized scanning illumination and oblique imaging over cleared, thick tissue sections in smooth motion, our approach achieves high-speed 3D image acquisition of an entire mouse brain within 2 hours, at a resolution capable of resolving synaptic spines. It is compatible with immunofluorescence labeling, enabling flexible cell-type specific brain mapping, and is readily scalable for large biological samples such as primate brain.

scholarly journals Light-Sheet Microscopy in Neuroscience

2019 ◽  
Vol 42 (1) ◽  
pp. 295-313 ◽  
Author(s):  
Elizabeth M.C. Hillman ◽  
Venkatakaushik Voleti ◽  
Wenze Li ◽  
Hang Yu
Keyword(s):  
High Speed ◽  
Light Sheet ◽  
Mammalian Brain ◽  
Diverse Range ◽  
Volumetric Imaging ◽  
Two Photon Microscopy ◽  
Light Sheet Microscopy ◽  
Noise Benefits ◽  
Longitudinal Imaging

Light-sheet microscopy is an imaging approach that offers unique advantages for a diverse range of neuroscience applications. Unlike point-scanning techniques such as confocal and two-photon microscopy, light-sheet microscopes illuminate an entire plane of tissue, while imaging this plane onto a camera. Although early implementations of light sheet were optimized for longitudinal imaging of embryonic development in small specimens, emerging implementations are capable of capturing light-sheet images in freely moving, unconstrained specimens and even the intact in vivo mammalian brain. Meanwhile, the unique photobleaching and signal-to-noise benefits afforded by light-sheet microscopy's parallelized detection deliver the ability to perform volumetric imaging at much higher speeds than can be achieved using point scanning. This review describes the basic principles and evolution of light-sheet microscopy, followed by perspectives on emerging applications and opportunities for both imaging large, cleared, and expanded neural tissues and high-speed, functional imaging in vivo.

scholarly journals Cortical Column and Whole Brain Imaging of Neural Circuits with Molecular Contrast and Nanoscale Resolution

2018 ◽  
Author(s):  
Ruixuan Gao ◽  
Shoh M. Asano ◽  
Srigokul Upadhyayula ◽  
Pisarev Igor ◽  
Daniel E. Milkie ◽  
...  
Keyword(s):  
Neural Development ◽  
High Speed ◽  
Large Scale ◽  
Neural Circuits ◽  
Light Sheet ◽  
Synaptic Proteins ◽  
Light Sheet Microscopy ◽  
Mouse Cortex ◽  
Molecular Contrast ◽  
The Brain

AbstractOptical and electron microscopy have made tremendous inroads in understanding the complexity of the brain, but the former offers insufficient resolution to reveal subcellular details and the latter lacks the throughput and molecular contrast to visualize specific molecular constituents over mm-scale or larger dimensions. We combined expansion microscopy and lattice light sheet microscopy to image the nanoscale spatial relationships between proteins across the thickness of the mouse cortex or the entire Drosophila brain, including synaptic proteins at dendritic spines, myelination along axons, and presynaptic densities at dopaminergic neurons in every fly neuropil domain. The technology should enable statistically rich, large scale studies of neural development, sexual dimorphism, degree of stereotypy, and structural correlations to behavior or neural activity, all with molecular contrast.One Sentence SummaryCombined expansion and lattice light sheet microscopy enables high speed, nanoscale molecular imaging of neural circuits over large volumes.

scholarly journals Performance trade-offs for single- and dual-objective light-sheet microscope designs

2020 ◽  
Author(s):  
Kevin W. Bishop ◽  
Adam K. Glaser ◽  
Jonathan T.C. Liu
Keyword(s):  
High Speed ◽  
Numerical Aperture ◽  
Light Sheet ◽  
Model Organisms ◽  
Design Parameters ◽  
Volumetric Imaging ◽  
Light Sheet Microscopy ◽  
Trade Offs ◽  
Moderate Resolution ◽  
Working Distance

AbstractLight-sheet microscopy (LSM) has emerged as a powerful tool for high-speed volumetric imaging of live model organisms and large optically cleared specimens. When designing cleared-tissue LSM systems with certain desired imaging specifications (e.g. resolution, contrast, and working distance), various design parameters must be taken into consideration. In order to elucidate some of the key design trade-offs for LSM systems, we present a diffraction-based analysis of single- and dual-objective LSM configurations where Gaussian illumination is utilized. Specifically, we analyze the effects of the illumination and collection numerical aperture (NA), as well as their crossing angle, on spatial resolution and contrast. Assuming an open-top light-sheet (OTLS) architecture, we constrain these parameters based on fundamental geometric considerations as well as those imposed by currently available microscope objectives. In addition to revealing the performance tradeoffs of various single- and dual-objective LSM configurations, our analysis showcases the potential advantages of a novel, non-orthogonal dual-objective (NODO) architecture, especially for moderate-resolution imaging applications (collection NA of 0.5 to 0.8).

scholarly journals High-speed panoramic light-sheet microscopy reveals global endodermal cell dynamics

2013 ◽  
Vol 4 (1) ◽  
Author(s):  
Benjamin Schmid ◽  
Gopi Shah ◽  
Nico Scherf ◽  
Michael Weber ◽  
Konstantin Thierbach ◽  
...  
Keyword(s):  
High Speed ◽  
Light Sheet ◽  
Endodermal Cell ◽  
Cell Dynamics ◽  
Light Sheet Microscopy

Deep-learning on-chip light-sheet microscopy enabling video-rate volumetric imaging of dynamic biological specimens

Lab on a Chip ◽  
2021 ◽  
Author(s):  
Xiaopeng Chen ◽  
Junyu Ping ◽  
Yixuan Sun ◽  
Chengqiang Yi ◽  
Sijian Liu ◽  
...  
Keyword(s):  
Deep Learning ◽  
Light Scattering ◽  
Light Sheet ◽  
High Contrast ◽  
Spatiotemporal Resolution ◽  
Volumetric Imaging ◽  
Light Sheet Microscopy ◽  
High Spatiotemporal Resolution ◽  
On Chip

Volumetric imaging of dynamic signals in a large, moving, and light-scattering specimen is extremely challenging, owing to the requirement on high spatiotemporal resolution and difficulty in obtaining high-contrast signals. Here...

scholarly journals Diagonally Scanned Light-Sheet Microscopy for Fast Volumetric Imaging of Adherent Cells

2016 ◽  
Vol 110 (6) ◽  
pp. 1456-1465 ◽  
Author(s):  
Kevin M. Dean ◽  
Philippe Roudot ◽  
Carlos R. Reis ◽  
Erik S. Welf ◽  
Marcel Mettlen ◽  
...  
Keyword(s):  
Light Sheet ◽  
Adherent Cells ◽  
Volumetric Imaging ◽  
Light Sheet Microscopy

Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy

2012 ◽  
Vol 9 (7) ◽  
pp. 755-763 ◽  
Author(s):  
Raju Tomer ◽  
Khaled Khairy ◽  
Fernando Amat ◽  
Philipp J Keller
Keyword(s):  
High Speed ◽  
Light Sheet ◽  
High Speed Imaging ◽  
Light Sheet Microscopy

Efficient processing and analysis of large-scale light-sheet microscopy data

2015 ◽  
Vol 10 (11) ◽  
pp. 1679-1696 ◽  
Author(s):  
Fernando Amat ◽  
Burkhard Höckendorf ◽  
Yinan Wan ◽  
William C Lemon ◽  
Katie McDole ◽  
...  
Keyword(s):  
Large Scale ◽  
Light Sheet ◽  
Light Sheet Microscopy ◽  
Efficient Processing ◽  
Microscopy Data

scholarly journals A hybrid open-top light-sheet microscope for multi-scale imaging of cleared tissues

2021 ◽  
Author(s):  
Adam Glaser ◽  
Kevin Bishop ◽  
Lindsey Barner ◽  
Etsuo Susaki ◽  
Shimpei Kubota ◽  
...  
Keyword(s):  
Hybrid System ◽  
High Throughput ◽  
Light Sheet ◽  
Refractive Indices ◽  
Volumetric Imaging ◽  
Tissue Clearing ◽  
Sample Geometry ◽  
Multi Scale ◽  
Light Sheet Microscopy ◽  
User Friendly

Abstract Light-sheet microscopy has emerged as the preferred means for high-throughput volumetric imaging of cleared tissues. However, there is a need for a user-friendly system that can address imaging applications with varied requirements in terms of resolution (mesoscopic to sub-micrometer), sample geometry (size, shape, and number), and compatibility with tissue-clearing protocols and sample holders of various refractive indices. We present a ‘hybrid’ system that combines a novel non-orthogonal dual-objective and conventional (orthogonal) open-top light-sheet architecture for versatile multi-scale volumetric imaging.

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