N. A. Rumyantseva, D. M. Golofeeva, I. E. Vishnyakov, A. D. Vedyaykin
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引用次数: 0
摘要
摘要 膨胀显微镜(ExM)是一种样品制备技术,可通过样品的物理膨胀来改善生物结构的可视化。这种方法与传统的光学显微镜结合使用,无需使用超分辨率显微镜所需的复杂技术设备,就能以更高的分辨率观察生物结构。与超分辨率显微镜的方法不同,膨胀显微镜无法克服衍射极限;不过,观察到的效果可被视为等同于空间分辨率的提高。这种方法相对简单,对所用显微镜的要求也不高,因此近年来膨胀显微镜已成为一种相当流行的观察各种生物结构的方法。本文介绍了利用膨胀显微镜观察 DNA 和大肠杆菌细胞中 FtsZ 蛋白在 SOS 反应过程中形成的结构。研究结果证实了之前获得的数据,即处于 SOS 反应状态的细胞中的 FtsZ 蛋白分布不均匀。这项工作中使用的利用膨胀显微镜方法观察大肠杆菌细胞的方案今后可用于研究其他细胞(包括细菌和真核细胞)的内部结构。
Visualization of Single Escherichia coli Cells in the State of SOS Response using Expansion Microscopy
Abstract
Expansion microscopy (ExM) is a sample preparation technique which allows to achieve improved visualization of biological structures based on the physical expansion of the sample. This method is used in combination with traditional light microscopy and allows to achieve visualization of biological structures with higher resolution, without the use of complex technical devices typical for super-resolution microscopy. Unlike the methods of super-resolution microscopy, expansion microscopy does not make it possible to overcome the diffraction limit; however, the observed effect can be considered equal to an increase in the spatial resolution. The relative simplicity of the method and low requirements for the microscope used, have made expansion microscopy a fairly popular method to visualize various biological structures recently. This paper describes the use of expansion microscopy to visualize DNA and structures formed by the FtsZ protein in Escherichia coli cells during the SOS response. The results of the work confirm the previously obtained data that the FtsZ protein in cells in the state of the SOS response is unevenly distributed. The protocol used in this work for visualization of E. coli cells using the expansion microscopy method can be used in the future to study the internal structures of other cells, both bacterial and eukaryotic.
期刊介绍:
The journal publishes papers on vast aspects of cell research, including morphology, biochemistry, biophysics, genetics, molecular biology, immunology. The journal accepts original experimental studies, theoretical articles suggesting novel principles and approaches, presentations of new hypotheses, reviews highlighting major developments in cell biology, discussions. The main objective of the journal is to provide a competent representation and integration of research made on cells (animal and plant cells, both in vivo and in cell culture) offering insight into the structure and functions of live cells as a whole. Characteristically, the journal publishes articles on biology of free-living and parasitic protists, which, unlike Metazoa, are eukaryotic organisms at the cellular level of organization.