Liquid-phase scanning electron microscopy for single membrane protein imaging

Wang, Li; Li, Changshuo; Li, Jintao; Zhang, Xiaofei; Li, Xiaochen; Cui, Yiran; Xia, Yang; Zhang, Yinqi; Mao, Shengcheng; Yuan, Jun; Wang, Sheng; Han, Xiaodong

doi:10.1016/j.bbrc.2021.12.081

Cited by 3 publications

(6 citation statements)

References 46 publications

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“…In particular, considering the large size of protein assemblies and eukaryotic cells, liquid-phase SEM equipped with analytical capability might serve as a complementary approach with TEM. By labeling with Au NPs or fluorescent quantum dots and binding them to features of interest, the spatial position, density, and arrangement of these electron-dense labels can provide significant insights into the corresponding positions and functions of the targeted intracellular structures , and resolving the dynamic structures of proteins and living cells . Similarly, a recent study reported the connection of mass spectrometry to ETEM (Figure g), realizing the real-time detection of gas species from oxidation of CNTs .…”

Section: Discussionmentioning

confidence: 99%

“…By labeling with Au NPs or fluorescent quantum dots and binding them to features of interest, the spatial position, density, and arrangement of these electron-dense labels can provide significant insights into the corresponding positions and functions of the targeted intracellular structures 676,677 and resolving the dynamic structures of proteins and living cells. 540 Similarly, a recent study reported the connection of mass spectrometry to ETEM (Figure 50g), realizing the real-time detection of gas species from oxidation of CNTs. 450 Such design may open more opportunities to the deeper investigation into reactions such as alcohol oxidation and the Fischer−Tropsch process, and the combination of mass spectrometry (and even NMR, high-performance liquid chromatography) with liquid-phase EM can be expected.…”

Section: Opportunities In In Situ (S)temmentioning

confidence: 90%

“…By sandwiching Au-labeled epidermal growth factor receptors (EGFRs) (on triple-negative breast cancer (TNBC) cells) between two SiN x microchips, the localization, distribution, and configuration of labeled membrane proteins can be precisely recognized. 540 One can also directly grow live cells on Si microchips. 541 Maintained in a saturated water vapor atmosphere, the hydrated cell is covered with a thin layer of water, mimicking the liquid environment.…”

Section: Liquid-phase Sem (And Stem-in-sem) Characterization Of the M...mentioning

confidence: 99%

“…To extend to other solvents, liquid-phase SEM utilizes SiN x microchips to achieve sealing against a high vacuum and benefits from a configuration of STEM-in-SEM or low-voltage STEM. By sandwiching Au-labeled epidermal growth factor receptors (EGFRs) (on triple-negative breast cancer (TNBC) cells) between two SiN x microchips, the localization, distribution, and configuration of labeled membrane proteins can be precisely recognized . One can also directly grow live cells on Si microchips .…”

Section: In Situ Characterization Of Soft Nanomaterials’ Full Life Cy...mentioning

confidence: 99%

“…Obtaining chemical information can also be achieved at a decently high resolution using SEM and ETEM. For example, given SEM's compatibility with surface characterization and large/thick samples, wider application of liquid-phase SEM is expected, such as tracking the 3D assembly process of NPs, characterizing reactions at the electrode−electrolyte interface (Figure 50a−f), 676 and resolving the dynamic structures of proteins and living cells, 540 among others. In particular, considering the large size of protein assemblies and eukaryotic cells, liquid-phase SEM equipped with analytical capability might serve as a complementary approach with TEM.…”

Section: Opportunities In In Situ (S)temmentioning

confidence: 99%

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Electron Microscopy Studies of Soft Nanomaterials

et al. 2023

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This review highlights recent efforts on applying electron microscopy (EM) to soft (including biological) nanomaterials. We will show how developments of both the hardware and software of EM have enabled new insights into the formation, assembly, and functioning (e.g., energy conversion and storage, phonon/photon modulation) of these materials by providing shape, size, phase, structural, and chemical information at the nanometer or higher spatial resolution. Specifically, we first discuss standard real-space two-dimensional imaging and analytical techniques which are offered conveniently by microscopes without special holders or advanced beam technology. The discussion is then extended to recent advancements, including visualizing three-dimensional morphology of soft nanomaterials using electron tomography and its variations, identifying local structure and strain by electron diffraction, and recording motions and transformation by in situ EM. On these advancements, we cover state-of-the-art technologies designed for overcoming the technical barriers for EM to characterize soft materials as well as representative application examples. The even more recent integration of machine learning and its impacts on EM are also discussed in detail. With our perspectives of future opportunities offered at the end, we expect this review to inspire and stimulate more efforts in developing and utilizing EM-based characterization methods for soft nanomaterials at the atomic to nanometer length scales in academic research and industrial applications.

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Section: Discussionmentioning

confidence: 99%

Section: Opportunities In In Situ (S)temmentioning

confidence: 90%

Section: Liquid-phase Sem (And Stem-in-sem) Characterization Of the M...mentioning

confidence: 99%

Section: In Situ Characterization Of Soft Nanomaterials’ Full Life Cy...mentioning

confidence: 99%

Section: Opportunities In In Situ (S)temmentioning

confidence: 99%

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Electron Microscopy Studies of Soft Nanomaterials

et al. 2023

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In situ liquid cell SEM observation of dynamic processes of Au nanoparticles

Chen

Tan

Cheng

et al. 2023

Microscopy Res & Technique

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In situ liquid cell electron microscopy (LC‐EM) is a powerful platform for real time nanoscale imaging of liquid systems. In situ liquid cell scanning electron microscopy (LC‐SEM) as a relatively low cost and potentially more convenient characterization method, has not been as widely used as compared to in situ liquid cell transmission electron microscopy (LC‐TEM). This paper reports a real time high resolution and comprehensive characterization of Au nanoparticles (NPs) and nanoparticle clusters (NPCs), which are surface‐decorated with cetyltrimethylammonium bromide (CTAB), in an oleic acid (OA) emulsion system with LC‐SEM. Single NP resolution images are routinely collected with both secondary electron (SE) and backscattered electron (BSE) imaging modes, with different SEM systems. Energy dispersive spectroscopy (EDS) mapping data clearly demonstrates the single particle level chemical element distributions, particle stacking structure, as well as the preferred distribution of OA molecules on the Au particle surfaces. Moreover, both liquid droplet growth and particle motions are observed with LC‐SEM, among which, ways for faster tracking the single particle level dynamic motion behavior of Au NPs and NPCs are explored. We expect that our work will bring new insight of high resolution and fast analysis in a broad range of materials in liquid with LC‐SEM.

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Atomic force microscopy: A nanobiotechnology for cellular research

Guan

2022

Nano TransMed

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Nanobiotechnology such as atomic force microscopy (AFM) has a great application in various regimes of cell biology, offering an excellent avenue to study cellular nanotopography, nanomechanics, and nanointeraction. AFM nanotopography can provide a high resolution of nano-architectures of different cells. AFM nanomechanics have shed new light on characterizing mechanical properties of cellular structures and biological materials as well as monitoring the physiopathological processes. AFM nanointeraction measurement helps the understanding of the molecular interaction forces at a nanoscale.

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Liquid-phase scanning electron microscopy for single membrane protein imaging

Cited by 3 publications

References 46 publications

Electron Microscopy Studies of Soft Nanomaterials

Electron Microscopy Studies of Soft Nanomaterials

In situ liquid cell SEM observation of dynamic processes of Au nanoparticles

Atomic force microscopy: A nanobiotechnology for cellular research

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