Scanning ion conductance microscopy of isolated metaphase chromosomes in a liquid environment
Scanning ion conductance microscopy of isolated metaphase chromosomes in a liquid environment
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液体环境中分离的中期染色体的扫描离子电导显微镜
DOI：
10.1007/s10577-021-09659-0
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发表时间：
2021
影响因子：
2.6
通讯作者：
F. Iwata
中科院分区：
文献类型：
--
作者：
T. Ushiki;K. Ishizaki;Y. Mizutani;M. Nakajima;F. Iwata
扫描探针显微镜（SPM）使用探针尖扫描样品表面以获取样品表面特征的信息。在各种类型的SPM中，原子力显微镜（AFM）已广泛应用于包括染色体在内的生物样品的成像。扫描离子电导显微镜（SICM）也被引入到可视化生物样品的表面结构中，因为它可以通过检测通过移液器开口的离子流来获得液体条件下的“无接触”地形图像。然而，我们最近注意到，SICM很难对染色体进行一致成像。本文对离子流在样品表面的行为进行了精确的研究，以期获得比目前更一致的离体猕猴中期染色体的SICM图像。本研究表明，在移液管电极上应用正电位可以获得染色体的地形图像，而应用负电位则无法成像。研究了移液器接近染色体时的离子电流与尖端样品距离的关系。电流-电压（I-V）曲线进一步为我们提供了染色体成像过程中离子电流行为的准确解释。这些数据进一步与HeLa细胞的SICM成像数据进行比较。我们的研究结果表明，在正常的Dulbecco磷酸盐缓冲盐水中，染色体是带电的，净电荷是强烈的负电荷。结果表明，离子浓度对SICM成像染色体有重要影响。
Scanning probe microscopy (SPM) uses a probing tip which scans over a sample surface for obtaining information on the sample surface characteristics. Among various types of SPM, atomic force microscopy (AFM) has been widely applied to imaging of biological samples including chromosomes. Scanning ion conductance microscopy (SICM) has been also introduced for visualizing the surface structure of biological samples because it can obtain “contact-free” topographic images in liquid conditions by detecting ion current flow through a pipette opening. However, we recently noticed that the consistent imaging of chromosomes is difficult by SICM. In this paper, the behaviors of the ion current on the sample surfaces were precisely investigated for obtaining SICM images of isolated muntjac metaphase chromosomes more consistently than at present. The present study revealed that application of positive potential to the pipette electrode was acceptable for obtaining the topographic image of chromosomes, while application of negative potential failed in imaging. The approach curves were then studied for analyzing the relationship between the ion current and the tip sample distance when the pipette is approaching chromosomes. The current-voltage (I-V) curve further provided us the accurate interpretation of the ion current behavior during chromosome imaging. These data were further compared with those for SICM imaging of HeLa cells. Our findings indicated that chromosomes are electrically charged and the net charge is strongly negative in normal Dulbecco’s phosphate buffered saline. We finally showed that the ion concentration of the bath electrolyte is important for imaging chromosomes by SICM.
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影响因子：
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作者：
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DOI：
--
发表时间：
2021
期刊：
影响因子：
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作者：
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