Max Planck Institute of Cell Biology and Genetics, 01309 Dresden, Germany.
Max Planck Institute of Cell Biology and Genetics, 01309 Dresden, Germany.
Methods. 2018 May 1;140-141:188-197. doi: 10.1016/j.ymeth.2017.12.005. Epub 2017 Dec 16.
Quantifying molecular dynamics of cell membrane constituents is required to understand organization and function of biological membranes. Because of its complex structure unambiguous interpretation of molecular membrane dynamics requires high spatial and temporal resolution measurements. In this paper, we provide a comprehensive description of circle scanning fluorescence correlation spectroscopy and its combination with stimulated emission depletion microscopy (CS-STED-FCS). This method allows quantification of sub-diffusion processes and direct mapping of heterogeneities in membranes with high spatiotemporal resolution. We show how to use model membranes to calibrate and test the technique and how to apply it in the context of living cells to quantify membrane dynamics with high spatiotemporal resolution and good statistics.
为了理解生物膜的组织和功能，需要定量研究细胞膜成分的分子动力学。由于其复杂的结构，明确解释分子膜动力学需要高空间和时间分辨率的测量。在本文中，我们提供了圆扫描荧光相关光谱及其与受激发射损耗显微镜（CS-STED-FCS）结合的综合描述。该方法允许定量亚扩散过程，并以高时空分辨率直接绘制膜中的非均质性。我们展示了如何使用模型膜来校准和测试该技术，以及如何将其应用于活细胞中，以高时空分辨率和良好的统计数据来量化膜动力学。
