High-precision thickness control of ice layer on CVD grown bilayer graphene for cryo-TEM

Kato, Ryuichi; Hatano, Yuri; Kasahata, Naoki; Sato, Chikara; Suenaga, Kazu; Hasegawa, Masataka

doi:10.1016/j.carbon.2020.01.010

Cited by 12 publications

(6 citation statements)

References 40 publications

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“…The clean graphene film with promising hydrophilicity would also have potential for the applications in cryo-EM imaging by enabling the formation of uniform thin ice layers and minimization of the background signal, where graphene would function as the supporting film. ,, To evaluate the impact of surface contamination on the spatial distribution of water on graphene surfaces, cryoenvironmental scanning EM (ESEM) and cryo-TEM characterizations were conducted on the suspended graphene membrane (Figure a and Figure S12a). Before the cryo-ESEM imaging, the suspended graphene samples were first exposed to stable water vapors for 10 s and then immediately put into the liquid nitrogen to prepare the vitrified cryosamples.…”

Section: Resultsmentioning

confidence: 99%

Hydrophilic, Clean Graphene for Cell Culture and Cryo-EM Imaging

et al. 2021

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The wettability of graphene is critical for numerous applications but is very sensitive to its surface cleanness. Herein, by clarifying the impact of intrinsic contamination, i.e., amorphous carbon, which is formed on the graphene surface during the high-temperature chemical vapor deposition (CVD) process, the hydrophilic nature of clean graphene grown on single-crystal Cu(111) substrate was confirmed by both experimental and theoretical studies, with an average water contact angle of ∼23°. Furthermore, the wettability of as-transferred graphene was proven to be highly dependent on its intrinsic cleanness, because of which the hydrophilic, clean graphene exhibited improved performance when utilized for cell culture and cryoelectron microscopy imaging. This work not only validates the intrinsic hydrophilic nature of graphene but also provides a new insight in developing advanced bioapplications using CVDgrown clean graphene films.

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Section: Resultsmentioning

confidence: 99%

Hydrophilic, Clean Graphene for Cell Culture and Cryo-EM Imaging

et al. 2021

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“…Finally, owing to its high level of crystallinity, graphene has a highly ordered and stable structure, and its interference with the images can be easily filtered out. In this context, H3N2 influenza viruses were imaged by liquid cell TEM, while tobacco mosaic virus was observed by conventional TEM, cryo-TEM, , and low-energy electron holography, resolving the helical arrangement of the outer protein shell of the tobacco mosaic virus.…”

Section: Interactions Between Viruses and 2d Materialsmentioning

confidence: 99%

Two-Dimensional Material-Based Biosensors for Virus Detection

2020

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Viral infections are one of the major causes of mortality and economic losses worldwide. Consequently, efficient virus detection methods are crucial to determine the infection prevalence. However, most detection methods face challenges related to false-negative or falsepositive results, long response times, high costs, and/or the need for specialized equipment and staff. Such issues can be overcome by access to low-cost and fast response point-of-care detection systems, and two-dimensional materials (2DMs) can play a critical role in this regard. Indeed, the unique and tunable physicochemical properties of 2DMs provide many advantages for developing biosensors for viral infections with high sensitivity and selectivity. Fast, accurate, and reliable detection, even at early infection stages by the virus, can be potentially enabled by highly accessible surface interactions between the 2DMs and the analytes. High selectivity can be obtained by functionalization of the 2DMs with antibodies, nucleic acids, proteins, peptides or aptamers, allowing for specific binding to a particular virus, viral fingerprints or proteins released by the host organism. Multiplexed detection and discrimination between different virus strains are also feasible. In this review, we present a comprehensive overview of the major advances of 2DMbased biosensors for the detection of viruses. We describe the main factors governing the efficient interactions between viruses and 2DMs, making them ideal candidates for the detection of viral infections. We also critically detail their advantages and drawbacks, providing insights for the development of future biosensors for virus detection. Lastly, we provide suggestions to stimulate research in the fast expanding field of 2DMs that could help in designing advanced systems for preventing virus-related pandemics.

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“…Samples (3.5 μL) were applied onto a UV-O3-treated lacey supported grid and then blotted for 1 s to remove an excess solution before rapid vitrification by plunge-freezing the grids into liquid ethane. [30] Grids were transferred to a Gatan cryo stage and imaged using JEM-1400 (JEOL Ltd., Tokyo, Japan) operating at 120 kV equipped with a Gatan 914 cryo-holder (Gatan Inc., Pleasanton, CA, USA). TEM images were recorded and investigated using a Veleta CCD camera and RA-DIUS (Olympus-SiS, Münster, Germany).…”

Section: Methodsmentioning

confidence: 99%

Conjugation‐Free Multilamellar Protein‐Lipid Hybrid Vesicles for Multifaceted Immune Responses

Koo

Jin

Kim

et al. 2021

Adv Healthcare Materials

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Various lipid-based nanocarriers have been developed for the co-delivery of protein antigens with immunological adjuvants. However, their in vivo potency in vaccine delivery is limited by structural instability, which causes off-target delivery and low cross-presentation efficacies. Recent works employ covalent cross-linking to stabilize the lipid nanostructures, though the immunogenicity and side effects of chemically modified protein antigens and lipids can cause a long-lasting safety issue. Here robust "conjugation-free" multilamellar protein antigen-lipid hybrid nanovesicles (MPLVs) are introduced through the antigen-mediated self-assembly of unilamellar lipid vesicles for the co-delivery of protein antigens and immunologic adjuvants. The nanocarriers coated with monophosphoryl lipid A and hyaluronic acids elicit highly increase antigen-specific immune responses in vitro and in vivo. The MPLVs increase the generation of immunological surface markers and cytokines in mouse-derived bone-marrow dendritic cells compared to soluble antigens with adjuvants. Besides, the vaccination of mice with the MPLVs significantly increase the production of anti-antigen antibody and interferon-gamma via the activation of CD4 + and CD8 + T cells, respectively. These findings suggest that MPLVs can serve as a promising nanovaccine delivery platform for efficient antigen cross-presentation through the efficient co-delivery of protein antigens with adjuvants.

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High-precision thickness control of ice layer on CVD grown bilayer graphene for cryo-TEM

Cited by 12 publications

References 40 publications

Hydrophilic, Clean Graphene for Cell Culture and Cryo-EM Imaging

Hydrophilic, Clean Graphene for Cell Culture and Cryo-EM Imaging

Two-Dimensional Material-Based Biosensors for Virus Detection

Conjugation‐Free Multilamellar Protein‐Lipid Hybrid Vesicles for Multifaceted Immune Responses

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