Scanning ion-conductance microscopy with a double-barreled nanopipette for topographic imaging of charged chromosomes

IF 1.8 4区 工程技术
Microscopy Pub Date : 2021-10-01 DOI:10.1093/jmicro/dfab009
Futoshi Iwata;Tatsuru Shirasawa;Yusuke Mizutani;Tatsuo Ushiki
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引用次数: 4

Abstract

Scanning ion conductance microscopy (SICM) is useful for imaging soft and fragile biological samples in liquids because it probes the samples’ surface topography by detecting ion currents under non-contact and force-free conditions. SICM acquires the surface topographical height by detecting the ion current reduction that occurs when an electrolyte-filled glass nanopipette approaches the sample surface. However, most biological materials have electrically charged surfaces in liquid environments, which sometimes affect the behavior of the ion currents detected by SICM and, especially, make topography measurements difficult. For measuring such charged samples, we propose a novel imaging method that uses a double-barrel nanopipette as an SICM probe. The ion current between the two apertures of the nanopipette desensitizes the surface charge effect on imaging. In this study, metaphase chromosomes of Indian muntjac were imaged by this technique because, owing to their strongly negatively charged surfaces in phosphate-buffered saline, it is difficult to obtain the topography of the chromosomes by the conventional SICM with a single-aperture nanopipette. Using the proposed method with a double-barrel nanopipette, the surfaces of the chromosomes were successfully measured, without any surface charge confounder. Since the detailed imaging of sample topography can be performed in physiological liquid conditions regardless of the sample charge, it is expected to be used for analyzing the high-order structure of chromosomes in relation to their dynamic changes in the cell division.
用双管纳米移液管扫描离子电导显微镜对带电染色体进行形貌成像
扫描离子电导显微镜(SICM)可用于对液体中柔软易碎的生物样品进行成像,因为它通过在非接触和无力条件下检测离子电流来探测样品的表面形貌。SICM通过检测填充电解质的玻璃纳米移液管接近样品表面时发生的离子电流减少来获取表面形貌高度。然而,大多数生物材料在液体环境中具有带电表面,这有时会影响SICM检测到的离子电流的行为,尤其是使形貌测量变得困难。为了测量这种带电样品,我们提出了一种新的成像方法,使用双管纳米移液管作为SICM探针。纳米移液管的两个孔之间的离子电流使成像中的表面电荷效应不敏感。在本研究中,通过该技术对印度魔芋的中期染色体进行了成像,因为它们在磷酸盐缓冲盐水中的表面带有强烈的负电荷,很难用单孔纳米移液管通过传统的SICM获得染色体的形貌。使用所提出的双管纳米移液管的方法,成功地测量了染色体的表面,没有任何表面电荷混淆。由于样品形貌的详细成像可以在生理液体条件下进行,而与样品电荷无关,因此有望用于分析染色体的高阶结构及其在细胞分裂中的动态变化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Microscopy
Microscopy 工程技术-显微镜技术
自引率
11.10%
发文量
0
审稿时长
>12 weeks
期刊介绍: Microscopy, previously Journal of Electron Microscopy, promotes research combined with any type of microscopy techniques, applied in life and material sciences. Microscopy is the official journal of the Japanese Society of Microscopy.
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