Robust ultraclean atomically thin membranes for atomic-resolution electron microscopy

Zheng, Liming; Chen, Yanan; Li, Ning; Zhang, Jincan; Liu, Nan; Liu, Junjie; Dang, Wenhui; Deng, Bing; Li, Yanbin; Gao, Xiaoyin; Tan, Congwei; Yang, Zi; Xu, Shipu; Wang, Mingzhan; Yang, Hao; Sun, Luzhao; Cui, Yi; Wei, Xiaoding; Gao, Peng; Wang, Hongwei; Peng, Hailin

doi:10.1038/s41467-020-14359-0

Cited by 49 publications

(42 citation statements)

References 46 publications

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“…Basically, it is a cleaning technique with water-methanol mixture and centrifugation of the solution for two-three times to isolate large GO flakes, where the majority of GO flakes are 20–70 μm. The previous study by Zheng et al showed that ~20–50 μm GO monolayer functionalized-GO monolayer separations [ 45 , 46 ] could be isolated using low pressure Chemical Vapor Deposition (LPCVD) system successfully. Additionally, NTA modified graphene oxide monolayer was successfully isolated for TEM grids preparation [ 44 ], and the overall size of the GO monolayer is between ~16 and 37 μm.…”

Section: Resultsmentioning

confidence: 99%

“…Our visual inspection, manual screening of individual holes of entire grids (5 grids are screened manually) and statistical analysis suggest that GO monolayer isolated by H 2 O-Methanol is appropriate to coat individual square (50 μm × 50 μm) of holey carbon grids ( Figure 5 ). However, all the GO coated grids were used to determine the cryo-EM structure of proteasome [ 24 , 45 ], GroEL [ 44 ], and apoferritin [ 47 ]. Nevertheless, none of them were able to characterize a 4.6Å map using only ~7800 single particle projections using GO coated holey grids.…”

Section: Resultsmentioning

confidence: 99%

“…Over the previous decade, several methods, like exfoliation, chemical treatment (highly acidic or basic solutions) and organic solvent treatment, have been developed to isolate GO monolayer [ 10 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 ]. Recently, few researchers deployed functionalized graphene oxide to prepare the grids and cryo-EM sample [ 4 , 23 , 24 , 42 , 43 , 44 , 45 , 46 ], although overall GO isolation procedure is expensive, tedious and time consuming. However, Graphene-based materials or graphene treated with water/methanol mixture have not been rigorously tested for preparing grids for room temperature TEM or cryo-EM study.…”

Section: Introductionmentioning

confidence: 99%

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Simplified Approach for Preparing Graphene Oxide TEM Grids for Stained and Vitrified Biomolecules

2021

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In this manuscript, we report the application of graphene oxide (GO) in the preparation of cryo-electron microscopy (cryo-EM) and transmission electron microscopy (TEM) grids. We treated GO with water and organic solvents, such as, methanol, ethanol and isopropanol separately to isolate significantly large GO monolayer flake to fabricate the grids for cryo-EM and TEM study. We implemented a simplified approach to isolate flakes of GO monolayer for constructing the TEM grids, independent of expensive heavy equipment (Langmuir–Blodgett trough, glow-discharge system, carbon-evaporator or plasma-cleaner or peristaltic pumps). We employed confocal microscopy, SEM and TEM to characterize the flake size, stability and transparency of the GO monolayer and atomic force microscopy (AFM) to probe the depth of GO coated grids. Additionally, GO grids are visualized at cryogenic condition for suitability of GO monolayer for cryo-EM study. In addition, GO-Met-H2O grids reduce the effect of preferred orientation of biological macromolecules within the amorphous ice. The power-spectrum and contrast-transfer-function unequivocally suggest that GO-Met-H2O fabricated holey grids have excellent potential for application in high-resolution structural characterization of biomolecules. Furthermore, only 200 movies and ~8000 70S ribosome particles are selected on GO-coated grids for cryo-EM reconstruction to achieve high-resolution structure.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Simplified Approach for Preparing Graphene Oxide TEM Grids for Stained and Vitrified Biomolecules

2021

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“…[ 18 ] As shown in Figure , the blooming field of 2D materials comprises different structure types and compositions, such as elemental analogs of graphene (EAs), MXenes, TMDs, covalent organic frameworks (COFs), and metal organic frameworks (MOFs), among which EAs, MXenes, and TMDs are introduced preferentially here. [ 19,20 ] Similar to graphene, EAs contain one kind of element, mostly from Groups IV or V of the periodic table. The ones composed of Group IV elements, such as silicene, germanene, and stanene, have the same electronic configurations as graphene.…”

Section: Methodsmentioning

confidence: 99%

In Situ and Operando Characterizations of 2D Materials in Electrochemical Energy Storage Devices

et al. 2021

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The urgent need of modern society for portable energy‐consuming devices has boosted the development of high‐power supercapacitors and high‐energy batteries. Major prerequisites for augmenting wider practical applications are advanced electrode materials and deeper insights into the underlying electrochemical processes. Owing to their unique physicochemical properties, 2D materials, such as graphene, transition metal carbides, nitrides, and dichalcogenides, hold special promise. Characterizing their behavior under real operating conditions (operando) or at the site of operation (in situ) without disassembling the device helps uncover essential kinetic information, which may, otherwise, be lost. By identifying both harmful and beneficial mechanisms, these in situ and operando characterization techniques allow researchers to alleviate critical issues in existing materials and develop new materials with enhanced properties. Herein, a brief introduction including the preparation and the electrochemical energy storage application of 2D materials is first presented. The main concern, thereby, is the influence of preparation methods on the resulting electrode structure and electrochemical performance. Then, the electrochemical mechanisms underlying the operation of supercapacitors and lithium batteries are discussed in detail. Finally, the perspective and future direction of applying the in situ and operando techniques to model 2D materials in revealing some important processes are highlighted.

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“…This partially dsRNA was then mixed with L protein in a 1.1:1 molar ratio at 4°C for 3 hours and the complex LRNA was ultimately resolved on Superdex 200 10/300 GL in a gel filtration buffer (50 mM BTP pH 8.5, 300 mM NaCl). Aliquots of 4 μL purified RVFV LRNA protein at a concentration of~0.4 mg/ml were applied to glow-discharged holey carbon Cu grids (Quantifoil R1.2/1.3) and 4 μL purified RVFV Lapo protein at a concentration of~0.264 mg/ml were applied to glow-discharged homemade graphene grids 44 . Then these two kinds of grids were flash-cooled in liquid ethane using an FEI Vitrobot Mark IV.…”

Section: Cryo-em Sample Preparation and Data Collectionmentioning

confidence: 99%

Structural insights into Rift Valley fever virus replication machinery

Chen¹,

Wang²,

Wang³

et al. 2021

Preprint

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Rift Valley fever virus (RVFV) belongs to the order Bunyavirales and is the type species of genus Phlebovirus, which accounts for over 50% of family Phenuiviridae species. RNA-dependent RNA polymerase (L protein) is responsible for facilitating the replication and transcription of the virus. We report two cryo-EM RVFV L protein structures at 3.6 Å and 3.8 Å resolution in the presence and absence of RNA, respectively. In this first L protein structure of genus Phlebovirus, viral RNA induces considerable conformational changes of the polymerase. The RVFV L protein priming loop is distinctly different from those of other L proteins and undergoes large movements related to its replication elongation role. Structural and biochemical analyses indicate that a single template can initiate RNA replication, which is notably enhanced by 5’ viral RNA. These findings advance our understanding of RNA synthesis mechanism and provide a basis for antiviral inhibitor development.

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Robust ultraclean atomically thin membranes for atomic-resolution electron microscopy

Cited by 49 publications

References 46 publications

Simplified Approach for Preparing Graphene Oxide TEM Grids for Stained and Vitrified Biomolecules

Simplified Approach for Preparing Graphene Oxide TEM Grids for Stained and Vitrified Biomolecules

In Situ and Operando Characterizations of 2D Materials in Electrochemical Energy Storage Devices

Structural insights into Rift Valley fever virus replication machinery

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