Nanoarchitecture and dynamics of the mouse enteric glycocalyx examined by freeze-etching electron tomography and intravital microscopy

AbstractThe glycocalyx is a highly hydrated, glycoprotein-rich coat shrouding many eukaryotic and prokaryotic cells. The intestinal epithelial glycocalyx, comprising glycosylated transmembrane mucins, is part of the primary host-microbe interface and is essential for nutrient absorption. Its disruption has been implicated in numerous gastrointestinal diseases. Yet, due to challenges in preserving and visualizing its native organization, glycocalyx structure-function relationships remain unclear. Here, we characterize the nanoarchitecture of the murine enteric glycocalyx using freeze-etching and electron tomography. Micrometer-long mucin filaments emerge from microvillar-tips and, through zigzagged lateral interactions form a three-dimensional columnar network with a 30 nm mesh. Filament-termini converge into globular structures ~30 nm apart that are liquid-crystalline packed within a single plane. Finally, we assess glycocalyx deformability and porosity using intravital microscopy. We argue that the columnar network architecture and the liquid-crystalline packing of the filament termini allow the glycocalyx to function as a deformable size-exclusion filter of luminal contents.

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Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010005161x ◽

1974 ◽

Vol 32 ◽

pp. 320-321

Author(s):

J. P. Revel

Keyword(s):

Developmental Stages ◽

Three Dimensional ◽

Cell Movement ◽

Cellular Interactions ◽

Serial Sections ◽

Cell Sheets ◽

Freeze Etching ◽

Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

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Characterization of photosystem II with the atomic-force microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100130365 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1146-1147

Author(s):

Kathleen M. Marr ◽

Mary K. Lyon

Keyword(s):

Photosystem Ii ◽

Atomic Force Microscope ◽

Three Dimensional ◽

Biologically Active ◽

Atomic Force ◽

Freeze Etching ◽

Image Averaging ◽

Oxygen Evolving ◽

Averaging Techniques

Photosystem II (PSII) is different from all other reaction centers in that it splits water to evolve oxygen and hydrogen ions. This unique ability to evolve oxygen is partly due to three oxygen evolving polypeptides (OEPs) associated with the PSII complex. Freeze etching on grana derived insideout membranes revealed that the OEPs contribute to the observed tetrameric nature of the PSIl particle; when the OEPs are removed, a distinct dimer emerges. Thus, the surface of the PSII complex changes dramatically upon removal of these polypeptides. The atomic force microscope (AFM) is ideal for examining surface topography. The instrument provides a topographical view of individual PSII complexes, giving relatively high resolution three-dimensional information without image averaging techniques. In addition, the use of a fluid cell allows a biologically active sample to be maintained under fully hydrated and physiologically buffered conditions. The OEPs associated with PSII may be sequentially removed, thereby changing the surface of the complex by one polypeptide at a time.

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Practical electron tomography

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100140087 ◽

1995 ◽

Vol 53 ◽

pp. 742-743

Author(s):

C.L. Woodcock

Keyword(s):

Electron Tomography ◽

Tomographic Reconstruction ◽

Three Dimensional ◽

3D Shape ◽

Computational Resources ◽

Shape Of Particles

Despite the potential of the technique, electron tomography has yet to be widely used by biologists. This is in part related to the rather daunting list of equipment and expertise that are required. Thanks to continuing advances in theory and instrumentation, tomography is now more feasible for the non-specialist. One barrier that has essentially disappeared is the expense of computational resources. In view of this progress, it is time to give more attention to practical issues that need to be considered when embarking on a tomographic project. The following recommendations and comments are derived from experience gained during two long-term collaborative projects.Tomographic reconstruction results in a three dimensional description of an individual EM specimen, most commonly a section, and is therefore applicable to problems in which ultrastructural details within the thickness of the specimen are obscured in single micrographs. Information that can be recovered using tomography includes the 3D shape of particles, and the arrangement and dispostion of overlapping fibrous and membranous structures.

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Three-Dimensional reconstruction of isolated centrosomes by automated electron tomography

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100169286 ◽

1994 ◽

Vol 52 ◽

pp. 310-311

Author(s):

M.B. Braunfeld ◽

M. Moritz ◽

B.M. Alberts ◽

J.W. Sedat ◽

D.A. Agard

Keyword(s):

Electron Tomography ◽

Three Dimensional ◽

Vital Role ◽

Complex Nature ◽

Animal Cells ◽

Microtubule Organizing Center ◽

Nucleation Sites ◽

Dimensional Reconstruction ◽

Organizing Center ◽

Resolution Model

In animal cells, the centrosome functions as the primary microtubule organizing center (MTOC). As such the centrosome plays a vital role in determining a cell's shape, migration, and perhaps most importantly, its division. Despite the obvious importance of this organelle little is known about centrosomal regulation, duplication, or how it nucleates microtubules. Furthermore, no high resolution model for centrosomal structure exists.We have used automated electron tomography, and reconstruction techniques in an attempt to better understand the complex nature of the centrosome. Additionally we hope to identify nucleation sites for microtubule growth.Centrosomes were isolated from early Drosophila embryos. Briefly, after large organelles and debris from homogenized embryos were pelleted, the resulting supernatant was separated on a sucrose velocity gradient. Fractions were collected and assayed for centrosome-mediated microtubule -nucleating activity by incubating with fluorescently-labeled tubulin subunits. The resulting microtubule asters were then spun onto coverslips and viewed by fluorescence microscopy.

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An Update on Recent Advances in Cubosome: A Novel Drug Delivery System

Current Drug Metabolism ◽

10.2174/1389200221666210105121532 ◽

2021 ◽

Vol 21 ◽

Author(s):

Madhukar Garg ◽

Anju Goyal ◽

Sapna Kumari

Keyword(s):

Drug Delivery ◽

Drug Delivery System ◽

Lipid Bilayers ◽

Delivery System ◽

Liquid Crystalline ◽

Three Dimensional ◽

Biologically Active ◽

Potential Applications ◽

Health Related ◽

Novel Drug

: Cubosomes are highly stable nanostructured liquid crystalline dosage delivery form derived from amphiphilic lipids and polymer-based stabilizers converting it in a form of effective biocompatible carrier for the drug delivery. The delivery form comprised of bicontinuous lipid bilayers arranged in three dimensional honeycombs like structure provided with two internal aqueous channels for incorporation of number of biologically active ingredients. In contrast liposomes they provide large surface area for incorporation of different types of ingredients. Due to the distinct advantages of biocompatibility and thermodynamic stability, cubosomes have remained the first preference as method of choice in the sustained release, controlled release and targeted release dosage forms as new drug delivery system for the better release of the drugs. As lot of advancement in the new form of dosage form has bring the novel avenues in drug delivery mechanisms so it was matter of worth to compile the latest updates on the various aspects of mentioned therapeutic delivery system including its structure, routes of applications along with the potential applications to encapsulate variety drugs to serve health related benefits.

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Advances in Liquid Crystalline Epoxy Resins for High Thermal Conductivity

Polymers ◽

10.3390/polym13081302 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1302

Author(s):

Younggi Hong ◽

Munju Goh

Keyword(s):

Thermal Conductivity ◽

Network Structure ◽

Epoxy Resins ◽

Liquid Crystalline ◽

Random Network ◽

Three Dimensional ◽

Heat Insulator ◽

Liquid Crystalline Epoxy ◽

Thermally Conductive ◽

Substantial Interest

Epoxy resin (EP) is one of the most famous thermoset materials. In general, because EP has a three-dimensional random network, it possesses thermal properties similar to those of a typical heat insulator. Recently, there has been substantial interest in controlling the network structure of EP to create new functionalities. Indeed, the modified EP, represented as liquid crystalline epoxy (LCE), is considered promising for producing novel functionalities, which cannot be obtained from conventional EPs, by replacing the random network structure with an oriented one. In this paper, we review the current progress in the field of LCEs and their application to highly thermally conductive composite materials.

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Three-Dimensional Nanoparticle Distribution and Local Curvature of Heterogeneous Catalysts Revealed by Electron Tomography

The Journal of Physical Chemistry C ◽

10.1021/jp072441b ◽

2007 ◽

Vol 111 (31) ◽

pp. 11501-11505 ◽

Cited By ~ 55

Author(s):

Edmund P. W. Ward ◽

Timothy J. V. Yates ◽

José-Jesús Fernández ◽

David E. W. Vaughan ◽

Paul A. Midgley

Keyword(s):

Heterogeneous Catalysts ◽

Electron Tomography ◽

Three Dimensional ◽

Local Curvature ◽

Nanoparticle Distribution

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Capturing Enveloped Viruses on Affinity Grids for Downstream Cryo-Electron Microscopy Applications

Microscopy and Microanalysis ◽

10.1017/s1431927613013937 ◽

2013 ◽

Vol 20 (1) ◽

pp. 164-174 ◽

Cited By ~ 10

Author(s):

Gabriella Kiss ◽

Xuemin Chen ◽

Melinda A. Brindley ◽

Patricia Campbell ◽

Claudio L. Afonso ◽

...

Keyword(s):

Electron Microscopy ◽

Measles Virus ◽

Influenza A ◽

Electron Tomography ◽

Three Dimensional ◽

Nitrilotriacetic Acid ◽

Dimensional Structure ◽

Enveloped Viruses ◽

Cryo Electron Microscopy ◽

Human Immunodeficiency Virus Virus

AbstractElectron microscopy (EM), cryo-electron microscopy (cryo-EM), and cryo-electron tomography (cryo-ET) are essential techniques used for characterizing basic virus morphology and determining the three-dimensional structure of viruses. Enveloped viruses, which contain an outer lipoprotein coat, constitute the largest group of pathogenic viruses to humans. The purification of enveloped viruses from cell culture presents certain challenges. Specifically, the inclusion of host-membrane-derived vesicles, the complete destruction of the viruses, and the disruption of the internal architecture of individual virus particles. Here, we present a strategy for capturing enveloped viruses on affinity grids (AG) for use in both conventional EM and cryo-EM/ET applications. We examined the utility of AG for the selective capture of human immunodeficiency virus virus-like particles, influenza A, and measles virus. We applied nickel-nitrilotriacetic acid lipid layers in combination with molecular adaptors to selectively adhere the viruses to the AG surface. This further development of the AG method may prove essential for the gentle and selective purification of enveloped viruses directly onto EM grids for ultrastructural analyses.

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