Approach to map nanotopography of cell surface receptors.
Approach to map nanotopography of cell surface receptors.
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DOI：
10.1038/s42003-022-03152-y
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发表时间：
2022-03-09
影响因子：
5.9
通讯作者：
Cebecauer M
中科院分区：
文献类型：
--
作者：
Franke C;Chum T;Kvíčalová Z;Glatzová D;Gentsch GJ;Rodriguez A;Helmerich DA;Herdly L;Mavila H;Frank O;Brdička T;van de Linde S;Cebecauer M
细胞通过表面受体与环境进行交流，但纳米级受体组织与复杂细胞表面形态的关系尚不清楚。这主要是由于缺乏可获得的、可靠的和高分辨率的方法。在这里，我们提出了一种以纳米精度绘制细胞表面受体地形的方法。该方法包括用甘氨酸涂覆玻璃盖，这保留了精细的膜形态，同时允许固定细胞靠近光学表面定位。我们开发了一种先进的简化算法，用于分析双翼检测方案中获得的单分子定位数据。这些进步使蛋白质分布在皱褶质膜上的直接和定量映射具有接近各向同性的3D纳米分辨率。正如CD4和CD45受体成功证明的那样，所描述的工作流程是一种直接的定量技术，用于研究分子及其在分化后生细胞复杂表面纳米形态上的相互作用。一种基于超分辨率定位的方法可以绘制免疫细胞中的受体地形，从而可以定量研究分子及其在细胞复杂表面纳米形态上的相互作用。
Cells communicate with their environment via surface receptors, but nanoscopic receptor organization with respect to complex cell surface morphology remains unclear. This is mainly due to a lack of accessible, robust and high-resolution methods. Here, we present an approach for mapping the topography of receptors at the cell surface with nanometer precision. The method involves coating glass coverslips with glycine, which preserves the fine membrane morphology while allowing immobilized cells to be positioned close to the optical surface. We developed an advanced and simplified algorithm for the analysis of single-molecule localization data acquired in a biplane detection scheme. These advancements enable direct and quantitative mapping of protein distribution on ruffled plasma membranes with near isotropic 3D nanometer resolution. As demonstrated successfully for CD4 and CD45 receptors, the described workflow is a straightforward quantitative technique to study molecules and their interactions at the complex surface nanomorphology of differentiated metazoan cells. A super-resolution localisation-based method is shown to map receptor topography in immune cells, which allows quantitative study of molecules and their interactions at the complex surface nanomorphology of cells.
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影响因子：
5.5
作者：
Bernardino de la Serna J;Schütz GJ;Eggeling C;Cebecauer M
通讯作者：
Cebecauer M
影响因子：
7.3
作者：
Bálint Š;Lopes FB;Davis DM
通讯作者：
Davis DM
影响因子：
16.6
作者：
de Castro, Maria Angela Gomes;Wildhagen, Hanna;Opazo, Felipe
通讯作者：
Opazo, Felipe
影响因子：
16.6
作者：
Ehmann, Nadine;van de Linde, Sebastian;Alon, Amit;Ljaschenko, Dmitrij;Keung, Xi Zhen;Holm, Thorge;Rings, Annika;DiAntonio, Aaron;Hallermann, Stefan;Ashery, Uri;Heckmann, Manfred;Sauer, Markus;Kittel, Robert J.
通讯作者：
Kittel, Robert J.
影响因子：
30.5
作者：
Dustin ML;Davis SJ
通讯作者：
Davis SJ