Development of high-speed ion conductance microscopy

Watanabe, Shinji; Kitazawa, Satoko; Sun, Linhao; Kodera, Noriyuki; Ando, Toshio

doi:10.1063/1.5118360

Cited by 31 publications

(22 citation statements)

References 64 publications

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“…To further understand how genetic alterations cause the acquisition of malignant phenotypes, such as increased migration and invasiveness, we performed a nano-scale topography and mechanical property analysis using scanning ion conductance microscopy (SICM). An SICM analysis can create live images that simultaneously show topography and stiffness by contact-free scanning of the cell surface with a nanopipette that measures the ion migration through the pipette tip [ 25 ]. Notably, we observed a distinct topographic feature—a microridge structure—on the surface of AKT-common and AKF organoid cells, which actively fluctuate ( Fig.…”

Section: Main Subjectsmentioning

confidence: 99%

Genetic Alterations and Microenvironment that Drive Malignant Progression of Colorectal Cancer: Lessons from Mouse and Organoid Models

et al. 2022

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Comprehensive genome analyses have identified frequently mutated genes in human colorectal cancers (CRC). These include APC , KRAS , SMAD4 , TP53 , and FBXW7 . The biological functions of the respective gene products in cell proliferation and homeostasis have been intensively examined by in vitro experiments. However, how each gene mutation or combinations of specific mutations drive malignant progression of CRC in vivo has not been fully understood. Based on the genomic information, we generated mouse models that carry multiple mutations of CRC driver genes in various combinations, and we performed comprehensive histological analyses to link genetic alteration(s) and tumor phenotypes, including liver metastasis. In this review article, we summarize the phenotypes of the respective genetic models carrying major driver mutations and discuss a possible mechanism of mutations underlying malignant progression.

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Section: Main Subjectsmentioning

confidence: 99%

Genetic Alterations and Microenvironment that Drive Malignant Progression of Colorectal Cancer: Lessons from Mouse and Organoid Models

et al. 2022

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“…A recent study [40] revealed a significant contribution of electroosmotic flow and the SCD of the tip to the measured ion current when a large ion concentration gradient exists around a small aperture of the tip. To quantitatively analyze these contributions to the measured SICM data obtained under such conditions [41], information of the detailed tip geometry is indispensable.…”

Section: Importance Of Membrane On Tem Gridmentioning

confidence: 99%

Geometrical Characterization of Glass Nanopipettes with Sub-10 nm Pore Diameter by Transmission Electron Microscopy

et al. 2020

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Glass nanopipettes are widely used for various applications in nanosciences. In most of the applications, it is important to characterize their geometrical parameters, such as the aperture size and the inner cone angle at the tip region. For nanopipettes with sub-10 nm aperture and thin wall thickness, transmission electron microscopy (TEM) must be most instrumental in their precise geometrical measurement. However, this measurement has remained a challenge because heat generated by electron beam irradiation would largely deform sub-10-nm nanopipettes. Here we provide methods for preparing TEM specimens that do not cause deformation of such tiny nanopipettes.

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“…In this paper we report the adaptation and optimization of a wide-range x , y , z -nano-positioner design previously developed for high-speed SICM imaging 12 , 16 for HS-AFM scanning. The improved design extends the scanner’s acquisition bandwidth and permits high-fidelity, low-noise imaging at 0.5 fps (100 Hz line rate) over a 36 × 36 µm 2 area, corresponding to a high scan speed of 7.2 mm/s.…”

Section: Introductionmentioning

confidence: 99%

An ultra-wide scanner for large-area high-speed atomic force microscopy with megapixel resolution

Marchesi

Umeda

Komekawa

et al. 2021

Sci Rep

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High-speed atomic force microscopy (HS-AFM) is a powerful tool for visualizing the dynamics of individual biomolecules. However, in single-molecule HS-AFM imaging applications, x,y-scanner ranges are typically restricted to a few hundred nanometers, preventing overview observation of larger molecular assemblies, such as 2-dimensional protein crystal growth or fibrillar aggregation. Previous advances in scanner design using mechanical amplification of the piezo-driven x,y-positioning system have extended the size of HS-AFM image frames to several tens of micrometer, but these large scanners may suffer from mechanical instabilities at high scan speeds and only record images with limited pixel numbers and comparatively low lateral resolutions (> 20–100 nm/pixel), complicating single-molecule analysis. Thus, AFM systems able to image large sample areas at high speeds and with nanometer resolution have still been missing. Here, we describe a HS-AFM sample-scanner system able to record large topographic images (≤ 36 × 36 µm2) containing up to 16 megapixels, providing molecular resolution throughout the image frame. Despite its large size, the flexure-based scanner features a high resonance frequency (> 2 kHz) and delivers stable operation even at high scans speeds of up to 7.2 mm/s, minimizing the time required for recording megapixel scans. We furthermore demonstrate that operating this high-speed scanner in time-lapse mode can simultaneously identify areas of spontaneous 2-dimensional Annexin A5 crystal growth, resolve the angular orientation of large crystalline domains, and even detect rare crystal lattice defects, all without changing scan frame size or resolution. Dynamic processes first identified from overview scans can then be further imaged at increased frame rates in reduced scan areas after switching to conventional HS-AFM scanning. The added ability to collect large-area, high-resolution images of complex samples within biological-relevant time frames extends the capabilities of HS-AFM from single-molecule imaging to the study of large dynamic molecular arrays. Moreover, large-area HS-AFM scanning can generate detailed structural data sets from a single scan, aiding the quantitative analysis of structurally heterogenous samples, including cellular surfaces.

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Development of high-speed ion conductance microscopy

Cited by 31 publications

References 64 publications

Genetic Alterations and Microenvironment that Drive Malignant Progression of Colorectal Cancer: Lessons from Mouse and Organoid Models

Genetic Alterations and Microenvironment that Drive Malignant Progression of Colorectal Cancer: Lessons from Mouse and Organoid Models

Geometrical Characterization of Glass Nanopipettes with Sub-10 nm Pore Diameter by Transmission Electron Microscopy

An ultra-wide scanner for large-area high-speed atomic force microscopy with megapixel resolution

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