Large‐scale electron microscopic volume imaging of interfascicular oligodendrocytes in the mouse corpus callosum

Glia ◽  
2021 ◽  
Author(s):  
Tatsuhide Tanaka ◽  
Nobuhiko Ohno ◽  
Yasuyuki Osanai ◽  
Sei Saitoh ◽  
Truc Quynh Thai ◽  
...  
Keyword(s):  
Corpus Callosum ◽  
Large Scale ◽  
Electron Microscopic ◽  
Volume Imaging

scholarly journals Deciphering the role of trehalose in hindering antithrombin polymerization

2019 ◽  
Vol 39 (4) ◽  
Author(s):  
Asma Naseem ◽  
Mohammad Sazzad Khan ◽  
Hashim Ali ◽  
Irshad Ahmad ◽  
Mohamad Aman Jairajpuri
Keyword(s):  
Conformational Change ◽  
Large Scale ◽  
Guanidine Hydrochloride ◽  
Microscopic Analysis ◽  
Normal Activity ◽  
Significant Loss ◽  
Coagulation Cascade ◽  
Disulfonic Acid ◽  
Electron Microscopic ◽  
Polymer Formation

Abstract Serine protease inhibitors (serpins) family have a complex mechanism of inhibition that requires a large scale conformational change. Antithrombin (AT), a member of serpin superfamily serves as a key regulator of the blood coagulation cascade, deficiency of which leads to thrombosis. In recent years, a handful of studies have identified small compounds that retard serpin polymerization but abrogated the normal activity. Here, we screened small molecules to find potential leads that can reduce AT polymer formation. We identified simple sugar molecules that successfully blocked polymer formation without a significant loss of normal activity of AT under specific buffer and temperature conditions. Of these, trehalose proved to be most promising as it showed a marked decrease in the bead like polymeric structures of AT shown by electron microscopic analysis. A circular dichroism (CD) analysis indicated alteration in the secondary structure profile and an increased thermal stability of AT in the presence of trehalose. Guanidine hydrochloride (GdnHCl)-based unfolding studies of AT show the formation of a different intermediate in the presence of trehalose. A time-dependent fluorescence study using 1,1′-bi(4-anilino)naphthalene-5,5′-disulfonic acid (Bis-ANS) shows that trehalose affects the initial conformational change step in transition from native to polymer state through its binding to exposed hydrophobic residues on AT thus making AT less polymerogenic. In conclusion, trehalose holds promise by acting as an initial scaffold that can be modified to design similar compounds with polymer retarding propensity.

scholarly journals Conductive resins improve charging and resolution of acquired images in electron microscopic volume imaging

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Huy Bang Nguyen ◽  
Truc Quynh Thai ◽  
Sei Saitoh ◽  
Bao Wu ◽  
Yurika Saitoh ◽  
...  
Keyword(s):  
Electron Microscopic ◽  
Volume Imaging ◽  
Conductive Resins

scholarly journals Conneconomics: The Economics of Dense, Large-Scale, High-Resolution Neural Connectomics

2013 ◽  
Author(s):  
Adam H Marblestone ◽  
Evan R Daugharthy ◽  
Reza Kalhor ◽  
Ian D Peikon ◽  
Justus M Kebschull ◽  
...  
Keyword(s):  
Dna Sequencing ◽  
Mouse Brain ◽  
Large Scale ◽  
Fluorescent Microscopy ◽  
Electron Microscopic ◽  
Fixed Tissue ◽  
Isotropic Resolution ◽  
Brain Connectome ◽  
Electron Microscopic Image ◽  
Axon Tracing

We analyze the scaling and cost-performance characteristics of current and projected connectomics approaches, with reference to the potential implications of recent advances in diverse contributing fields. Three generalized strategies for dense connectivity mapping at the scale of whole mammalian brains are considered: electron microscopic axon tracing, optical imaging of combinatorial molecular markers at synapses, and bulk DNA sequencing of trans-synaptically exchanged nucleic acid barcode pairs. Due to advances in parallel-beam instrumentation, whole mouse brain electron microscopic image acquisition could cost less than $100 million, with total costs presently limited by image analysis to trace axons through large image stacks. Optical microscopy at 50 to 100 nm isotropic resolution could potentially read combinatorially multiplexed molecular information from individual synapses, which could indicate the identifies of the pre-synaptic and post-synaptic cells without relying on axon tracing. An optical approach to whole mouse brain connectomics may be achievable for less than $10 million and could be enabled by emerging technologies to sequence nucleic acids in-situ in fixed tissue via fluorescent microscopy. Novel strategies relying on bulk DNA sequencing, which would extract the connectome without direct imaging of the tissue, could produce a whole mouse brain connectome for $100k to $1 million or a mouse cortical connectome for $10k to $100k. Anticipated further reductions in the cost of DNA sequencing could lead to a $1000 mouse cortical connectome.

scholarly journals Large-scale characterization of the microvascular geometry in development and disease by tissue clearing and quantitative ultramicroscopy

2020 ◽  
pp. 0271678X2096185
Author(s):  
Artur Hahn ◽  
Julia Bode ◽  
Allen Alexander ◽  
Kianush Karimian-Jazi ◽  
Katharina Schregel ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Basal Ganglia ◽  
Corpus Callosum ◽  
Large Scale ◽  
Three Dimensional ◽  
Branch Length ◽  
Brain Regions ◽  
Computer Hardware ◽  
Vascular Networks ◽  
Tissue Clearing

Three-dimensional assessment of optically cleared, entire organs and organisms has recently become possible by tissue clearing and selective plane illumination microscopy (“ultramicroscopy”). Resulting datasets can be highly complex, encompass over a thousand images with millions of objects and data of several gigabytes per acquisition. This constitutes a major challenge for quantitative analysis. We have developed post-processing tools to quantify millions of microvessels and their distribution in three-dimensional datasets from ultramicroscopy and demonstrate the capabilities of our pipeline within entire mouse brains and embryos. Using our developed acquisition, segmentation, and analysis platform, we quantify physiological vascular networks in development and the healthy brain. We compare various geometric vessel parameters (e.g. vessel density, radius, tortuosity) in the embryonic spinal cord and brain as well as in different brain regions (basal ganglia, corpus callosum, cortex). White matter tract structures (corpus callosum, spinal cord) showed lower microvascular branch densities and longer vessel branch length compared to grey matter (cortex, basal ganglia). Furthermore, we assess tumor neoangiogenesis in a mouse glioma model to compare tumor core and tumor border. The developed methodology allows rapid quantification of three-dimensional datasets by semi-automated segmentation of fluorescently labeled objects with conventional computer hardware. Our approach can aid preclinical investigations and paves the way towards “quantitative ultramicroscopy”.

Three-dimensional imaging and physiology of live neurons and glia: Confocal light and correlative high-voltage Electron Microscopy

1993 ◽  
Vol 51 ◽  
pp. 162-163
Author(s):  
J.N. Turner ◽  
D.H. Szarowski ◽  
W. Shain ◽  
M. Davis-Cox ◽  
D.O. Carpenter ◽  
...  
Keyword(s):  
Large Scale ◽  
Culture Media ◽  
Shape Change ◽  
Complex Function ◽  
3D Structure ◽  
Three Dimensional ◽  
Electron Microscopic ◽  
High Voltage Electron Microscopy ◽  
Voltage Electron ◽  
And Function

Correlating physiologic measures with three-dimensional (3D) imaging at the light and electron microscopic levels is a powerful combination of methods for studying the structure and function of biological systems. Neurobiology is an ideal field for the application of these methods because neurons and glia have complex and extensive 3D structure, and their physiology is under intense study. Neurons, such as those studied here from Aplysia, can be more than 100 μm in diameter, and glia undergo large scale 3D shape change as a function of a number of physiologic parameters. The ability to accurately quantitate the 3D structure, volume and surface area of live neurons and glia is important to our understanding of the complex function of these cells.Neurons were isolated from the major ganglia of juvenile Aplysia Californica and glia were obtained from long term cultures of LRM 55 cells or as primary isolates from rats. Cultures were exposed to Dil dissolved in DMSO with or without 20% Pluronic F-127 and added to the culture media. The imaging instrument was an Olympus IMT-2 and a Bio-Rad MRC-600.

scholarly journals The corpus callosum as anatomical marker of intelligence? A critical examination in a large-scale developmental study

2017 ◽  
Vol 223 (1) ◽  
pp. 285-296 ◽  
Author(s):  
René Westerhausen ◽  
Charline-Marie Friesen ◽  
Darius A. Rohani ◽  
Stine K. Krogsrud ◽  
Christian K. Tamnes ◽  
...  
Keyword(s):  
Corpus Callosum ◽  
Large Scale ◽  
Critical Examination ◽  
Developmental Study

scholarly journals Nitzschia austriaca Hustedt: a characteristic diatom of Hungarian inland saline waters including a morphological comparison with the type material

Phytotaxa ◽  
2017 ◽  
Vol 308 (1) ◽  
pp. 54 ◽  
Author(s):  
ÉVA ÁCS ◽  
ANGÉLA FÖLDI ◽  
CARLOS E. WETZEL ◽  
CSABA F. VAD ◽  
KEVE T. KISS ◽  
...  
Keyword(s):  
Large Scale ◽  
Large Population ◽  
Taxonomic Status ◽  
Electron Microscopic Investigation ◽  
Interactive Effects ◽  
Type Material ◽  
Similar Species ◽  
Electron Microscopic ◽  
Morphological Comparison ◽  
Central European

A detailed scanning electron microscopic investigation was carried out to clarify the taxonomic status of a small sigmoid Nitzschia species, a potential indicator of Central European soda waters. We found this taxon to be one of the dominant epiphytic diatoms collected from sodic bomb crater ponds at Apaj (Hungary). The large population allowed for a morphometric comparison based on frustule ultrastructure with the type material of the most similar species, Nitzschia austriaca Hustedt that was originally described from a soda pan in the region. The results clearly demonstrated an overlap between the Apaj population and the type material of N. austriaca (based on NMDS analysis), therefore we argue that they represent the same taxon. An emended diagnosis of N. austriaca is given. Total suspended solids and total phosphorous proved to be the most important factors predicting the occurrence of the species, with possible interactive effects of conductivity and pH. We then expanded the distribution of the species by revisiting data originating from previous large-scale surveys targeting sodic habitats in Hungary. On the basis of our results, N. austriaca is a characteristic species for Central European soda waters, including the protected astatic soda pans, indicating their typical chemical and physical characteristics.

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