scholarly journals 3D-mapping of human lymph node and spleen reveals integrated neuronal, vascular, and ductal cell networks

2021 ◽  
Author(s):  
Seth Currlin ◽  
Harry Nick ◽  
Jerelyn Nick ◽  
Maigan Brusko ◽  
Hunter Hakimian ◽  
...  
Keyword(s):  
Lymph Node ◽  
3D Visualization ◽  
Sympathetic Innervation ◽  
Light Sheet ◽  
Elastic Membrane ◽  
Human Spleen ◽  
Neuronal Markers ◽  
Choline Acetyl Transferase ◽  
Light Sheet Microscopy ◽  
Cell Networks

As secondary lymphoid organs, the spleen and lymph node represent important hubs for both innate and adaptive immunity. Neuroanatomical and tracing data, largely derived from rodents, suggest that lymph nodes contain sensory and sympathetic innervation, whereas the spleen contains postganglionic sympathetic innervation, with conflicting views regarding the existence of cholinergic or vagal innervation. Herein, we map the neuronal, vascular, and sinus cell networks from human spleen and lymph node using highly multiplexed CODEX (CO-Detection by indEXing) and 3D light sheet microscopy of cleared tissues. These data demonstrate striking delineation of two distinct layers within the lymph node subcapsular sinus-the ceiling defined by Podoplanin expression and floor by LYVE1, which overlays the lymph node follicles. Within the lymph node interior, we observed a mesh-like vessel network innervated with GAP43 and beta3-tubulin. Dense perivascular innervation occurred in both tissues, including a subset of axonal processes expressing choline acetyl transferase (ChAT). Four neuronal markers (ChAT, PGP9.5, tyrosine hydroxylase, and beta3-tubulin) localized to the arterial tunica externa suggest expression in the nervi vasorum while GAP43 was expressed within the internal elastic membrane of arteries. These data represent highly novel 3D visualization of perivascular and periductal autonomic innervation within these two key human organs.

ClearVolume: open-source live 3D visualization for light-sheet microscopy

2015 ◽  
Vol 12 (6) ◽  
pp. 480-481 ◽  
Author(s):  
Loic A Royer ◽  
Martin Weigert ◽  
Ulrik Günther ◽  
Nicola Maghelli ◽  
Florian Jug ◽  
...  
Keyword(s):  
Open Source ◽  
3D Visualization ◽  
Light Sheet ◽  
Light Sheet Microscopy

scholarly journals 3D visualization and quantification of microvessels in the whole ischemic mouse brain using solvent-based clearing and light sheet microscopy

2017 ◽  
Vol 37 (10) ◽  
pp. 3355-3367 ◽  
Author(s):  
Erlen Lugo-Hernandez ◽  
Anthony Squire ◽  
Nina Hagemann ◽  
Alexandra Brenzel ◽  
Maryam Sardari ◽  
...  
Keyword(s):  
Mouse Brain ◽  
3D Visualization ◽  
Focal Cerebral Ischemia ◽  
Light Sheet ◽  
Artery Occlusion ◽  
Cerebral Microvessels ◽  
Light Sheet Microscopy ◽  
Low Viscosity ◽  
Ischemic Tissue ◽  
Cerebral Artery Occlusion

The visualization of cerebral microvessels is essential for understanding brain remodeling after stroke. Injection of dyes allows for the evaluation of perfused vessels, but has limitations related either to incomplete microvascular filling or leakage. In conventional histochemistry, the analysis of microvessels is limited to 2D structures, with apparent limitations regarding the interpretation of vascular circuits. Herein, we developed a straight-forward technique to visualize microvessels in the whole ischemic mouse brain, combining the injection of a fluorescent-labeled low viscosity hydrogel conjugate with 3D solvent clearing followed by automated light sheet microscopy. We performed transient middle cerebral artery occlusion in C57Bl/6j mice and acquired detailed 3D vasculature images from whole brains. Subsequent image processing, rendering and fitting of blood vessels to a filament model was employed to calculate vessel length density, resulting in 0.922 ± 0.176 m/mm3 in healthy tissue and 0.329 ± 0.131 m/mm3 in ischemic tissue. This analysis showed a marked loss of capillaries with a diameter ≤ 10 µm and a more moderate loss of microvessels in the range > 10 and ≤ 20 µm, whereas vessels > 20 µm were unaffected by focal cerebral ischemia. We propose that this protocol is highly suitable for studying microvascular injury and remodeling post-stroke.

scholarly journals Fast volumetric mapping of human brain slices

2020 ◽  
Author(s):  
Luca Pesce ◽  
Annunziatina Laurino ◽  
Vladislav Gavryusev ◽  
Giacomo Mazzamuto ◽  
Giuseppe Sancataldo ◽  
...  
Keyword(s):  
Human Brain ◽  
Brain Connectivity ◽  
Brain Slices ◽  
Light Sheet ◽  
Volumetric Imaging ◽  
Neuronal Markers ◽  
Light Sheet Microscopy ◽  
Inhibitory Population ◽  
Anatomical Information ◽  
The Brain

AbstractWe still lack a detailed map of the anatomical disposition of neurons in the human brain. A complete map would be an important step for deeply understanding the brain function, providing anatomical information useful to decipher the neuronal pattern in healthy and diseased conditions. Here, we present several important advances towards this goal, obtained by combining a new clearing method, advanced Light Sheet Microscopy and automated machinelearning based image analysis. We perform volumetric imaging of large sequentially stained human brain slices, labelled for two different neuronal markers NeuN and GAD67, discriminating the inhibitory population and reconstructing the brain connectivity.

scholarly journals Light Sheet Microscopy-Assisted 3D Analysis of SARS-CoV-2 Infection in the Respiratory Tract of the Ferret Model

Viruses ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 529
Author(s):  
Luca M. Zaeck ◽  
David Scheibner ◽  
Julia Sehl ◽  
Martin Müller ◽  
Donata Hoffmann ◽  
...  
Keyword(s):  
Respiratory Tract ◽  
3D Visualization ◽  
Three Dimensional ◽  
Light Sheet ◽  
3D Analysis ◽  
Respiratory Pathogens ◽  
Upper Respiratory Tract ◽  
Thin Sections ◽  
Light Sheet Microscopy

The visualization of viral pathogens in infected tissues is an invaluable tool to understand spatial virus distribution, localization, and cell tropism in vivo. Commonly, virus-infected tissues are analyzed using conventional immunohistochemistry in paraffin-embedded thin sections. Here, we demonstrate the utility of volumetric three-dimensional (3D) immunofluorescence imaging using tissue optical clearing and light sheet microscopy to investigate host–pathogen interactions of pandemic SARS-CoV-2 in ferrets at a mesoscopic scale. The superior spatial context of large, intact samples (>150 mm3) allowed detailed quantification of interrelated parameters like focus-to-focus distance or SARS-CoV-2-infected area, facilitating an in-depth description of SARS-CoV-2 infection foci. Accordingly, we could confirm a preferential infection of the ferret upper respiratory tract by SARS-CoV-2 and suggest clustering of infection foci in close proximity. Conclusively, we present a proof-of-concept study for investigating critically important respiratory pathogens in their spatial tissue morphology and demonstrate the first specific 3D visualization of SARS-CoV-2 infection.

scholarly journals 3D visualization of SARS-CoV-2 infection and receptor distribution in Syrian hamster lung lobes display distinct spatial arrangements

2021 ◽  
Author(s):  
Ilhan Tomris ◽  
Kim M. Bouwman ◽  
Youri Adolfs ◽  
Danny Noack ◽  
Roosmarijn van der Woude ◽  
...  
Keyword(s):  
Syrian Hamster ◽  
3D Visualization ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Light Sheet ◽  
Receptor Expression ◽  
Spike Protein ◽  
Microscopy Imaging ◽  
Light Sheet Microscopy ◽  
Lung Lobes

AbstractSARS-CoV-2 attaches to angiotensin-converting enzyme 2 (ACE2) to gain entry into cells after which the spike protein is cleaved by the transmembrane serine protease 2 (TMPRRS2) to facilitate viral-host membrane fusion. ACE2 and TMPRRS2 expression profiles have been analyzed at the genomic, transcriptomic, and single-cell RNAseq level, however, biologically relevant protein receptor organization in whole tissues is still poorly understood. To describe the organ-level architecture of receptor expression, related to the ability of ACE2 and TMPRRS2 to mediate infectivity, we performed a volumetric analysis of whole Syrian hamster lung lobes. Lung tissue of infected and control animals were stained using antibodies against ACE2 and TMPRRS2, combined with fluorescent spike protein and SARS-CoV-2 nucleoprotein staining. This was followed by light-sheet microscopy imaging to visualize expression patterns. The data demonstrates that infection is restricted to sites with both ACE2 and TMPRRS2, the latter is expressed in the primary and secondary bronchi whereas ACE2 is predominantly observed in the terminal bronchioles and alveoli. Conversely, infection completely overlaps at these sites where ACE2 and TMPRSS2 co-localize.

scholarly journals 3D flow field estimation and assessment for live cell fluorescence microscopy

2019 ◽  
Vol 36 (5) ◽  
pp. 1317-1325 ◽  
Author(s):  
Sandeep Manandhar ◽  
Patrick Bouthemy ◽  
Erik Welf ◽  
Gaudenz Danuser ◽  
Philippe Roudot ◽  
...  
Keyword(s):  
Moving Objects ◽  
3D Visualization ◽  
Automatic Measurement ◽  
Ground Truth ◽  
Light Sheet ◽  
Supplementary Information ◽  
Multidimensional Data ◽  
Spatiotemporal Resolution ◽  
Diverse Range ◽  
Light Sheet Microscopy

Abstract Motivation The revolution in light sheet microscopy enables the concurrent observation of thousands of dynamic processes, from single molecules to cellular organelles, with high spatiotemporal resolution. However, challenges in the interpretation of multidimensional data requires the fully automatic measurement of those motions to link local processes to cellular functions. This includes the design and the implementation of image processing pipelines able to deal with diverse motion types, and 3D visualization tools adapted to the human visual system. Results Here, we describe a new method for 3D motion estimation that addresses the aforementioned issues. We integrate 3D matching and variational approach to handle a diverse range of motion without any prior on the shape of moving objects. We compare different similarity measures to cope with intensity ambiguities and demonstrate the effectiveness of the Census signature for both stages. Additionally, we present two intuitive visualization approaches to adapt complex 3D measures into an interpretable 2D view, and a novel way to assess the quality of flow estimates in absence of ground truth. Availability and implementation https://team.inria.fr/serpico/data/3d-optical-flow-data/ Supplementary information Supplementary data are available at Bioinformatics online.

scholarly journals Achromatic terahertz Airy beam generation with dielectric metasurfaces

Nanophotonics ◽  
2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Qingqing Cheng ◽  
Juncheng Wang ◽  
Ling Ma ◽  
Zhixiong Shen ◽  
Jing Zhang ◽  
...  
Keyword(s):  
Biomedical Imaging ◽  
Frequency Dispersion ◽  
Light Sheet ◽  
Photonic Devices ◽  
Self Healing ◽  
Beam Focusing ◽  
Light Sheet Microscopy ◽  
Airy Beam ◽  
Potential Applications ◽  
Airy Beams

AbstractAiry beams exhibit intriguing properties such as nonspreading, self-bending, and self-healing and have attracted considerable recent interest because of their many potential applications in photonics, such as to beam focusing, light-sheet microscopy, and biomedical imaging. However, previous approaches to generate Airy beams using photonic structures have suffered from severe chromatic problems arising from strong frequency dispersion of the scatterers. Here, we design and fabricate a metasurface composed of silicon posts for the frequency range 0.4–0.8 THz in transmission mode, and we experimentally demonstrate achromatic Airy beams exhibiting autofocusing properties. We further show numerically that a generated achromatic Airy-beam-based metalens exhibits self-healing properties that are immune to scattering by particles and that it also possesses a larger depth of focus than a traditional metalens. Our results pave the way to the realization of flat photonic devices for applications to noninvasive biomedical imaging and light-sheet microscopy, and we provide a numerical demonstration of a device protocol.

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