scholarly journals 4D imaging of soft matter in liquid water

2021 ◽  
Author(s):  
Gabriele Marchello ◽  
Cesare De Pace ◽  
Silvia Acosta-Gutierrez ◽  
Ciro Lopez-Vazquez ◽  
Neil Wilkinson ◽  
...  

Water is a critical component for both function and structure of soft matter and it is what bestows the adjective soft. Imaging samples in liquid state is thus paramount to gathering structural and dynamical information of any soft materials. Herein we propose the use of liquid phase electron microscopy to expand ultrastructural analysis into dynamical investigations. We imaged two soft matter examples: a polymer micelle and a protein in liquid phase using transmission electron microscopy and demonstrate that the inherent Brownian motion associated with the liquid state can be exploited to gather three-dimensional information of the materials in their natural state. We call such an approach brownian tomography (BT). We combine BT with single particle analysis (Brownian particle analysis BPA) to image protein structures with a spatial resolution close that achievable using cryogenic TEM. We show that BPA allows sub-nanometer resolution of soft materials and enables to gather information on conformational changes, hydration dynamics, and the effect of thermal fluctuations.Abstract Figure

2018 ◽  
Vol 24 (S1) ◽  
pp. 248-249
Author(s):  
Joseph P. Patterson ◽  
Hanglong Wu ◽  
Alessandro Ianiro ◽  
Hao Su ◽  
A. Catarina C. Esteves ◽  
...  

2020 ◽  
Vol 32 (25) ◽  
pp. 2001582 ◽  
Author(s):  
Hanglong Wu ◽  
Heiner Friedrich ◽  
Joseph P. Patterson ◽  
Nico A. J. M. Sommerdijk ◽  
Niels Jonge

Author(s):  
Anshul Assaiya ◽  
Ananth Prasad Burada ◽  
Surbhi Dhingra ◽  
Janesh Kumar

Cryo-electron microscopy (CryoEM) has superseded X-ray crystallography and NMR to emerge as a popular and effective tool for structure determination in recent times. It has become indispensable for the characterization of large macromolecular assemblies, membrane proteins, or samples that are limited, conformationally heterogeneous, and recalcitrant to crystallization. Besides, it is the only tool capable of elucidating high-resolution structures of macromolecules and biological assemblies in situ. A state-of-the-art electron microscope operable at cryo-temperature helps preserve high-resolution details of the biological sample. The structures can be determined, either in isolation via single-particle analysis (SPA) or helical reconstruction, electron diffraction (ED) or within the cellular environment via cryo-electron tomography (cryoET). All the three streams of SPA, ED, and cryoET (along with subtomogram averaging) have undergone significant advancements in recent times. This has resulted in breaking the boundaries with respect to both the size of the macromolecules/assemblies whose structures could be determined along with the visualization of atomic details at resolutions unprecedented for cryoEM. In addition, the collection of larger datasets combined with the ability to sort and process multiple conformational states from the same sample are providing the much-needed link between the protein structures and their functions. In overview, these developments are helping scientists decipher the molecular mechanism of critical cellular processes, solve structures of macromolecules that were challenging targets for structure determination until now, propelling forward the fields of biology and biomedicine. Here, we summarize recent advances and key contributions of the three cryo-electron microscopy streams of SPA, ED, and cryoET.


Microscopy ◽  
2021 ◽  
Author(s):  
Yuri Nishino ◽  
Kanako Miyazaki ◽  
Mizuho Kaise ◽  
Atsuo Miyazawa

Abstract An emulsion, a type of soft matter, is complexed with at least two materials in the liquid state (e.g. water and oil). Emulsions are classified into two types: water in oil (W/O) and oil in water (O/W), depending on the strength of the emulsifier. The properties and behavior of emulsions are directly correlated with the size, number, localization and structure of the dispersed phases in the continuous phase. Therefore, an understanding of the microstructure comprising liquid-state emulsions is essential for producing and evaluating these emulsions. Generally, it is impossible for conventional electron microscopy to examine liquid specimens, such as emulsion. Recent advances in cryo-scanning electron microscopy (cryo-SEM) could allow us to visualize the microstructure of the emulsions in a frozen state. Immersion freezing in slush nitrogen has often been used for preparing the frozen samples of soft matters. This preparation could generate ice crystals, and they would deform the microstructure of specimens. High-pressure freezing contributes to the inhibition of ice-crystal formation and is commonly used for preparing frozen biological samples with high moisture content. In this study, we compared the microstructures of immersion-frozen and high-pressure frozen emulsions (O/W and W/O types, respectively). The cryo-SEM observations suggested that high-pressure freezing is more suitable for preserving the microstructure of emulsions than immersion freezing.


Author(s):  
J. R. Porter ◽  
J. I. Goldstein ◽  
D. B. Williams

Alloy scrap metal is increasingly being used in electric arc furnace (EAF) steelmaking and the alloying elements are also found in the resulting dust. A comprehensive characterization program of EAF dust has been undertaken in collaboration with the steel industry and AISI. Samples have been collected from the furnaces of 28 steel companies representing the broad spectrum of industry practice. The program aims to develop an understanding of the mechanisms of formation so that procedures to recover residual elements or recycle the dust can be established. The multi-phase, multi-component dust particles are amenable to individual particle analysis using modern analytical electron microscopy (AEM) methods.Particles are ultrasonically dispersed and subsequently supported on carbon coated formvar films on berylium grids for microscopy. The specimens require careful treatment to prevent agglomeration during preparation which occurs as a result of the combined effects of the fine particle size and particle magnetism. A number of approaches to inhibit agglomeration are currently being evaluated including dispersal in easily sublimable organic solids and size fractioning by centrifugation.


Author(s):  
C.D. Humphrey ◽  
T.L. Cromeans ◽  
E.H. Cook ◽  
D.W. Bradley

There is a variety of methods available for the rapid detection and identification of viruses by electron microscopy as described in several reviews. The predominant techniques are classified as direct electron microscopy (DEM), immune electron microscopy (IEM), liquid phase immune electron microscopy (LPIEM) and solid phase immune electron microscopy (SPIEM). Each technique has inherent strengths and weaknesses. However, in recent years, the most progress for identifying viruses has been realized by the utilization of SPIEM.


Author(s):  
F. Banhart ◽  
F.O. Phillipp ◽  
R. Bergmann ◽  
E. Czech ◽  
M. Konuma ◽  
...  

Defect-free silicon layers grown on insulators (SOI) are an essential component for future three-dimensional integration of semiconductor devices. Liquid phase epitaxy (LPE) has proved to be a powerful technique to grow high quality SOI structures for devices and for basic physical research. Electron microscopy is indispensable for the development of the growth technique and reveals many interesting structural properties of these materials. Transmission and scanning electron microscopy can be applied to study growth mechanisms, structural defects, and the morphology of Si and SOI layers grown from metallic solutions of various compositions.The treatment of the Si substrates prior to the epitaxial growth described here is wet chemical etching and plasma etching with NF3 ions. At a sample temperature of 20°C the ion etched surface appeared rough (Fig. 1). Plasma etching at a sample temperature of −125°C, however, yields smooth and clean Si surfaces, and, in addition, high anisotropy (small side etching) and selectivity (low etch rate of SiO2) as shown in Fig. 2.


2021 ◽  
Vol 7 (21) ◽  
pp. eabg5628
Author(s):  
Julien Bous ◽  
Hélène Orcel ◽  
Nicolas Floquet ◽  
Cédric Leyrat ◽  
Joséphine Lai-Kee-Him ◽  
...  

The antidiuretic hormone arginine-vasopressin (AVP) forms a signaling complex with the V2 receptor (V2R) and the Gs protein, promoting kidney water reabsorption. Molecular mechanisms underlying activation of this critical G protein–coupled receptor (GPCR) signaling system are still unknown. To fill this gap of knowledge, we report here the cryo–electron microscopy structure of the AVP-V2R-Gs complex. Single-particle analysis revealed the presence of three different states. The two best maps were combined with computational and nuclear magnetic resonance spectroscopy constraints to reconstruct two structures of the ternary complex. These structures differ in AVP and Gs binding modes. They reveal an original receptor-Gs interface in which the Gαs subunit penetrates deep into the active V2R. The structures help to explain how V2R R137H or R137L/C variants can lead to two severe genetic diseases. Our study provides important structural insights into the function of this clinically relevant GPCR signaling complex.


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