School of Biomedical Engineering, McMaster University, Hamilton, ON, Canada.
Department of Chemistry and Chemical Biology, McMaster University, Hamilton, ON, Canada.
Methods Mol Biol. 2022;2440:289-303. doi: 10.1007/978-1-0716-2051-9_17.
Raster image correlation spectroscopy (RICS) enables detecting and quantifying diffusion in live cells using standard commercial laser scanning confocal microscopes. Here, we describe a protocol based on RICS for measuring the lateral diffusion of two immunoreceptors within the plasma membrane of the macrophage cell line RAW 264.7. The sample images and measurements presented in this chapter were obtained from RICS analysis of Toll-like receptor 2 (TLR2) and cluster of differentiation 14 (CD14), which are transmembrane and membrane-anchored receptors, respectively. A step-by-step guideline is provided to acquire raster-scanned images and to extract the diffusion coefficients using RICS analysis.
光栅图像相关光谱学（RICS）可利用标准商用激光扫描共聚焦显微镜检测和量化活细胞中的扩散。本文介绍了一种基于 RICS 的方案，用于测量巨噬细胞系 RAW 264.7 细胞质膜中两种免疫受体的侧向扩散。本章中呈现的样本图像和测量结果来自 Toll 样受体 2（TLR2）和分化群 14（CD14）的 RICS 分析，它们分别是跨膜和膜锚定受体。本文提供了一个逐步指南，用于获取光栅扫描图像并使用 RICS 分析提取扩散系数。
