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使用 evanescent散射显微镜分析膜蛋白结合动力学和细胞粘附的无标签分析

Jiying Xu^1,2,3, Caixin Huang^1,4, Liangju Li^1,4

¹Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China. pfzhang@iccas.ac.cn.

|September 6, 2023

概括

发光散射显微镜 (ESM) 通过检测细胞粘附的变化来测量连接体与膜蛋白的相互作用. 这种无标签的方法使得实时运动分析和单个活细胞中的细胞活动跟踪成为可能.

科学领域:

生物物理学的生物物理.
细胞生物学细胞生物学
显微镜的使用方法

背景情况:

在单个活细胞中精确测量连接体与膜蛋白的相互作用,对于理解细胞过程和药物查至关重要.
现有的方法在提供实时,无标签的单细胞水平的动力分析方面面临挑战.

研究的目的:

开发一种用于测量单个活细胞中的联体膜蛋白相互作用的新方法.
使用 evanescent散射显微镜 (ESM) 进行无标签的分子结合的动力分析.
为了能够实时跟踪细胞粘附动态在带结合过程中.

主要方法:

使用 evanescent散射显微镜 (ESM) 来检测细胞粘附性质在连接体结合时的变化.
分析固定和活单细胞中的联结体蛋白动力学.
利用ESM的高时空分辨率实时跟踪细胞粘附部位,变形和迁移.

主要成果:

与膜蛋白结合的连接体改变了细胞粘附特性,独立于细胞微运动或连接体质量.
通过ESM,可以对蛋白质和与膜蛋白结合的小分子进行动态分析.
细胞粘附部位的实时跟踪提供了关于结合体结合过程中细胞活动的见解.

结论:

发光散射显微镜 (ESM) 提供了一种多功能和用户友好的,无标签的检测策略.
该方法以单细胞分辨率在现场分析生物系统中分子相互作用动态.
这种方法在药物查和理解涉及膜蛋白的细胞过程中具有潜在的应用.

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