Publisher：Electrochimica Acta  

The layer-by-layer (LbL) technique is a simple method employed to fabricate multi-layered thin films by alternately adsorbing anionic and cationic polyelectrolytes through the electrostatic interactions. In order to fabricate thin films with desirable structures and properties, it is crucial to understand how different fabricate conditions direct the thin film formation. In this study, we performed the In situ characterization of thin firm using scanning ion conductance microscopy (SICM), where the thickness as well as the surface roughness are evaluated for films with different layer numbers. Moreover, the effects of the pH and ionic strength on the fabrication of the thin films were investigated. SICM enabled the determination of the film thickness with a single layer level and the comprehension of the relationship between the surface morphology and fabrication conditions.

DOI： 10.1016/j.electacta.2023.143152

Scopus

Publishing type：Research paper (scientific journal)  

DOI： 10.1116/6.0002706

Web of Science

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Language：Japanese   Publisher：The Japanese Society of Microscopy  

<p>Visualization of the dynamics on the living cell surface is essential for understanding cellular functions such as migration, cancer invasion, and differentiation. In order to evaluate cellular dynamics, it is necessary to develop visualization techniques with nanoscale spatial resolution and non-invasion. Scanning ion conductance microscopy (SICM) is one of the most powerful microscopy techniques for obtaining fragile cell morphology under physiological conditions. In recent years, several research groups have improved the temporal resolution of SICM to visualize rapid cellular reactions. Here, we provide an overview of SICM, including the basic scanning mode and equipment, and recent researches on the development of high-speed SICM.</p>

DOI： 10.11410/kenbikyo.58.2_71

CiNii Research

Language：English   Publishing type：Research paper (scientific journal)  

Scanning ion conductance microscopy (SICM) is a promising tool for visualizing the dynamics of nanoscale cell surface topography. However, there are still no guidelines for fabricating nanopipettes with ideal shape consisting of small apertures and thin glass walls. Therefore, most of the SICM imaging has been at a standstill at the submicron scale. In this study, we established a simple and highly reproducible method for the fabrication of nanopipettes with sub-20 nm apertures. To validate the improvement in the spatial resolution, we performed time-lapse imaging of the formation and disappearance of endocytic pits as a model of nanoscale time-lapse topographic imaging. We have also successfully imaged the localization of the hot spot and the released extracellular vesicles. The nanopipette fabrication guidelines for the SICM nanoscale topographic imaging can be an essential tool for understanding cell-cell communication.

DOI： 10.1021/acs.analchem.3c01010

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Language：English   Publishing type：Research paper (scientific journal)  

Extracellular fine particles of various sizes and origins can be taken up by cells, affecting their function. Understanding the cellular uptake processes is crucial for understanding the cellular effects of these particles and the development of means to control their internalization. Although macropinocytosis is a possible pathway for the cellular uptake of particles larger than 0.2 μm, its contribution to cellular uptake in non-phagocytic cells is controversial. Using 3 μm polystyrene beads as a model particle, we aimed to assess the detailed modes of their cellular uptake by non-phagocytic HeLa cells. Cellular uptake was assessed using confocal, scanning electron, and scanning ion conductance microscopy analyses, together with inhibitor studies. Our results revealed that 3 μm beads were taken up by HeLa cells by an actin-, cholesterol-, and membrane protrusions-dependent noncanonical endocytic pathway, different from the canonical macropinocytic and phagocytic pathways. Our work provides a framework for studying the cellular uptake of extracellular fine particles.

DOI： 10.1039/d2bm01353c

Web of Science

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