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细胞内分子运输的对对应显微镜

Julissa Sanchez-Velasquez¹, Ashleigh Solano¹, Michelle A Digman²

¹School of Physics, Faculty of Science, University of Melbourne, Melbourne, Australia.

Nature protocols

|February 6, 2025

概括

双相关显微镜追踪活细胞中的光分子运动,使用时空强度波动. 这种方法量化了分子运输模式和与细胞内结构的相互作用.

科学领域:

细胞生物学细胞生物学
生物物理学的生物物理.
显微镜技术的使用方法

背景情况:

光相关谱学 (FCS) 分析分子动力学.
现有的方法可能缺乏复杂的细胞环境的空间分辨率.
追踪活细胞内的分子扩散需要先进的成像技术.

研究的目的:

介绍线扫描对相关显微镜 (LSCPM) 用于活细胞分子跟踪的协议.
为了使分子运输模式和细胞内屏障可访问性的量化.
用双通道获取来演示分子复合体形成的跟踪.

主要方法:

在共聚焦激光扫描显微镜 (CLSM) 上获取线扫描数据.
从光强度波动计算对相关函数 (pCF).
扩展到用于交叉对相关函数 (cross-pCF) 分析的双通道采集.

主要成果:

空间地图和统计量化的分子到达时间.
量化细胞内屏障的分子可访问性.
选择性跟踪的分子运输基于复杂的形成与连接体.

结论:

LSCPM为分析活细胞中远程分子扩散提供了强大的工具.
该协议允许对分子贩运和相互作用进行详细的调查.
这种方法增强了对复杂细胞架构中的分子动态的理解.

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