Light sheet microscopy with high spatial resolution based on polarized structured illumination beam modulated by the phase mask

2018 ◽  
Vol 81 (9) ◽  
pp. 959-965 ◽  
Author(s):  
Levan Nhu ◽  
Xiaona Wang ◽  
Yong Liu ◽  
Cuifang Kuang ◽  
Xu Liu
Keyword(s):  
Spatial Resolution ◽  
High Spatial Resolution ◽  
Light Sheet ◽  
Phase Mask ◽  
Structured Illumination ◽  
Light Sheet Microscopy

scholarly journals Simultaneous multiview capture and fusion improves spatial resolution in wide-field and light-sheet microscopy

Optica ◽  
2016 ◽  
Vol 3 (8) ◽  
pp. 897 ◽  
Author(s):  
Yicong Wu ◽  
Panagiotis Chandris ◽  
Peter W. Winter ◽  
Edward Y. Kim ◽  
Valentin Jaumouillé ◽  
...  
Keyword(s):  
Spatial Resolution ◽  
Light Sheet ◽  
Wide Field ◽  
Light Sheet Microscopy

scholarly journals Tiling light sheet selective plane illumination microscopy using discontinuous light sheets

2018 ◽  
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.

scholarly journals A polymer gel index-matched to water enables diverse applications in fluorescence microscopy

2020 ◽  
Author(s):  
Xiaofei Han ◽  
Yijun Su ◽  
Hamilton White ◽  
Kate M. O’Neill ◽  
Nicole Y. Morgan ◽  
...  
Keyword(s):  
Surface Passivation ◽  
Super Resolution ◽  
Light Sheet ◽  
Polymer Gel ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Neural Structure ◽  
Uv Curable ◽  
Isotropic Resolution ◽  
Light Sheet Microscopy

AbstractWe demonstrate diffraction-limited and super-resolution imaging through thick layers (tens-hundreds of microns) of BIO-133, a biocompatible, UV-curable, commercially available polymer with a refractive index (RI) matched to water. We show that cells can be directly grown on BIO-133 substrates without the need for surface passivation and use this capability to perform extended time-lapse volumetric imaging of cellular dynamics 1) at isotropic resolution using dual-view light-sheet microscopy, and 2) at super-resolution using instant structured illumination microscopy. BIO-133 also enables immobilization of 1) Drosophila tissue, allowing us to track membrane puncta in pioneer neurons, and 2) Caenorhabditis elegans, which allows us to image and inspect fine neural structure and to track pan-neuronal calcium activity over hundreds of volumes. Finally, BIO-133 is compatible with other microfluidic materials, enabling optical and chemical perturbation of immobilized samples, as we demonstrate by performing drug and optogenetic stimulation on cells and C. elegans.

scholarly journals Acoustic light-sheet microscopy

2021 ◽  
Author(s):  
Stefan Wunderl ◽  
Ayumu Ishijima ◽  
Etsuo Susaki ◽  
Zihui Xu ◽  
Hong Song ◽  
...  
Keyword(s):  
Light Propagation ◽  
High Spatial Resolution ◽  
Single Cells ◽  
Acoustic Pulse ◽  
Light Sheet ◽  
Wide Field ◽  
Axial Resolution ◽  
3D Objects ◽  
Light Sheet Microscopy ◽  
The Brain

Light-sheet imaging of 3D objects with high spatial resolution remains an open challenge because of the trade-off between field-of-view (FOV) and axial resolution originating from the diffraction of light. We developed acoustic light-sheet microscopy (acoustic LSM), which actively manipulates the light propagation inside a large sample to obtain wide-field microscopic images deep inside a target. By accurately coupling a light-sheet illumination pulse into a planar acoustic pulse, the light-sheet can be continuously guided over large distances. We imaged a fluorescence-labeled transparent mouse brain for the FOVs of 19.3 x 12.4 mm2 and 9.7 x 5.9 mm2 with resolved microstructures and single cells deep inside the brain. Acoustic LSM creates new opportunities for the application of light-sheet in the field of industry to basic science.

scholarly journals Minutes-timescale 3D isotropic imaging of entire organs at subcellular resolution by content-aware compressed-sensing light-sheet microscopy

2019 ◽  
Author(s):  
Chunyu Fang ◽  
Tingting Chu ◽  
Tingting Yu ◽  
Yujie Huang ◽  
Yusha Li ◽  
...  
Keyword(s):  
Compressed Sensing ◽  
Spatial Resolution ◽  
Light Sheet ◽  
System Level ◽  
Projection Neurons ◽  
Long Distance ◽  
Light Sheet Microscopy ◽  
Macro Scale ◽  
Subcellular Resolution ◽  
Content Aware

AbstractInstant 3D imaging of entire organs and organisms at cellular resolution is a recurring challenge in life science. Here we report on a computational light-sheet microscopy able to achieve minute-timescale mapping of entire macro-scale organs at high spatial resolution, thereby overcoming the throughput limit of current 3D microscopy implementations. Through combining a dual-side confocally-scanned Bessel light-sheet illumination which provides thinner-and-wider optical sectioning of deep tissues, with a content-aware compressed sensing (CACS) computation pipeline which further improves the contrast and resolution based on a single acquisition, our method yields 3D images with high, isotropic spatial resolution and rapid acquisition improved by two-orders of magnitude. We demonstrate the imaging of whole brain (∼400 mm3), entire gastrocnemius and tibialis muscles (∼200 mm3) of mouse at subcellular resolution (0.5-μm isovoxel) and ultra-high throughput of 5∼10 minutes per sample. Various system-level cellular analyses, such as mapping cell populations at different brain sub-regions, tracing long-distance projection neurons over the entire brain, and calculating neuromuscular junction occupancy across whole muscle, were also readily enabled by our method.

scholarly journals Multimodal Magnetic Resonance Histology and Light Sheet Imaging for Quantitative Neurogenetics of the Mouse

2021 ◽  
Author(s):  
G. Allan Johnson ◽  
Gary Cofer ◽  
James Cook ◽  
James Gee ◽  
Adam Hall ◽  
...  
Keyword(s):  
Magnetic Resonance ◽  
Spatial Resolution ◽  
Mouse Brain ◽  
Angular Resolution ◽  
Imaging Techniques ◽  
Brain Connectivity ◽  
Light Sheet ◽  
Whole Brain ◽  
Light Sheet Microscopy

Paul Lauterbur closed his seminal paper on MRI with the statement that "zeugmatographic (imaging) techniques should find many useful applications in studies of the internal structures, states and composition of microscopic objects" {Lauterbur, 1973 #967}. Magnetic resonance microscopy was subsequently demonstrated in 1986 by three groups{Aguayo, 1986 #968}{Eccles, 1986 #969}{Johnson, 1986 #970}. The application of MRI to the study of tissue structure, i.e. magnetic resonance histology (MRH) was suggested in 1993 {Johnson, 1993 #957}. MRH, while based on the same physical principals as MRI is something fundamentally different than the clinical exams which are typically limited to voxel dimensions of ~ 1 mm3. Preclinical imaging systems can acquire images with voxels ~ 1000 times smaller. The MR histology images presented here have been acquired at yet another factor of 1000 increase in spatial resolution. Figure S1 in the supplement shows a comparison of a state-of-the-art fractional anisotropy images of a C57 mouse brain in vivo @ 150 um resolution (voxel volume of 3.3 x10-3 mm3) with the atlas we have generated for this work at 15 um spatial resolution (voxel volume of 3.3 x 10-6 mm3). In previous work, we have demonstrated the utility of MR histology in neurogenetics at spatial/angular resolution of 45 um /46 angles {Wang N, 2020 #972}. At this spatial/angular resolution it is possible to map whole brain connectivity with high correspondence to retroviral tracers {Calabrese, 2015 #895}. But the MRH derived connectomes can be derived in less than a day where the retroviral tracer studies require months/years {Oh, 2014 #971}. The resolution index (angular samples/voxel volume) for this previous work was >500,000 {Johnson, 2018 #894}. Figure S2 shows a comparison between that previous work and the new atlas presented in this paper with a resolution index of 32 million. Light sheet microscopy (LSM) has undergone similar rapid evolution over the last 20 years. The invention of tissue clearing, advances in immuno histochemistry and development of selective plane illumination microscopy (SPIM) now make it possible to acquire whole mouse brain images at submicron spatial resolution with a vast array of cell specific markers{Ueda, 2020 #974}{Park, 2018 #953}{Murray, 2015 #952}{Gao, 2014 #973}. And these advantages can be realized in scan times of < 6hrs. The major limitation from these studies is the distortion in the tissue from dissection from the cranium, swelling from clearing and staining, and tissue damage from handling. We report here the merger of these two methods: 1. MRH with the brain in the skull to provide accurate geometry, cytoarchitectural measures using scalar imaging metrics and whole brain connectivity at 15 um isotropic spatial resolution with super resolution track density images @ 5 um isotropic resolution; 2. whole brain multichannel LSM @ 1.8x1.8x4.0 um; 3. a big image data infrastructure that enables label mapping from the atlas to the MR image, geometric correction to the light sheet data, label mapping to the light sheet volumes and quantitative extraction of regional cell density. These methods make it possible to generate a comprehensive collection of image derived phenotypes (IDP) of cells and circuits covering the whole mouse brain with throughput that can be scaled for quantitative neurogenetics.

scholarly journals Fast, multicolor 3-D imaging of brain organoids with a new single-objective two-photon virtual light-sheet microscope

2018 ◽  
Author(s):  
Irina Rakotoson ◽  
Brigitte Delhomme ◽  
Philippe Djian ◽  
Andreas Deeg ◽  
Maia Brunstein ◽  
...  
Keyword(s):  
Spatial Resolution ◽  
Three Dimensional ◽  
Numerical Aperture ◽  
Light Sheet ◽  
Confocal Microscope ◽  
Large Field ◽  
Excitation Light ◽  
Two Photon ◽  
Light Sheet Microscopy ◽  
Functional Features

ABSTRACTHuman inducible pluripotent stem cells (hiPSCs) hold a large potential for disease modeling. hiPSC-derived human astrocyte and neuronal cultures permit investigations of neural signaling pathways with subcellular resolution. Combinatorial cultures, and three-dimensional (3-D) embryonic bodies enlarge the scope of investigations to multi-cellular phenomena. A the highest level of complexity, brain organoids that – in many aspects – recapitulate anatomical and functional features of the developing brain permit the study of developmental and morphological aspects of human disease. An ideal microscope for 3-D tissue imaging at these different scales would combine features from both confocal laser-scanning and light-sheet microscopes: a micrometric optical sectioning capacity and sub-micrometric spatial resolution, a large field of view and high frame rate, and a low degree of invasiveness, i.e., ideally, a better photon efficiency than that of a confocal microscope. In the present work, we describe such an instrument that belongs to the class of two-photon (2P) light-sheet microsocpes. Its particularity is that – unlike existing two- or three-lens designs – it is using a single, low-magnification, high-numerical aperture objective for the generation and scanning of a virtual light sheet. The microscope builds on a modified Nipkow-Petran spinning-disk scheme for achieving wide-field excitation. However, unlike the common Yokogawa design that uses a tandem disk, our concept combines micro lenses, dichroic mirrors and detection pinholes on a single disk. This design, advantageous for 2P excitation circumvents problems arising with the tandem disk from the large wavelength-difference between the infrared excitation light and visible fluorescence. 2P fluorescence excited in by the light sheet is collected by the same objective and imaged onto a fast sCMOS camera. We demonstrate three-dimensional imaging of TO-PRO3-stained embryonic bodies and of brain organoids, under control conditions and after rapid (partial) transparisation with triethanolamine and /ormamide (RTF) and compare the performance of our instrument to that of a confocal microscope having a similar numerical aperture. 2P-virtual light-sheet microscopy permits one order of magnitude faster imaging, affords less photobleaching and permits better depth penetration than a confocal microscope with similar spatial resolution.

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