质膜蛋白动力学的调控及其研究方法

罗鹏云, 钱虹萍, 刘艳, 徐昌文, 崔亚宁

Regulation of Plasma Membrane Protein Dynamics and Its Research Methods

Pengyun Luo, Hongping Qian, Yan Liu, Changwen Xu, Yaning Cui

图2 质膜蛋白动力学技术 (A) 由折射到全内反射(total internal reflection, TIR): 1束光在2种不同的介质中传播, 介质的折射率分别为n1和n2, 入射角为α, 由Snell定律可知, 光会发生折射进入另一种介质, 且在另一种介质中的传播角度与原介质不同, 当入射角α1等于甚至大于临界角α2时, 光就不会进入到较小折射率的介质中, 而是完全被反射到原来的介质中, 即发生全内反射; (B) 在荧光相关光谱法(FCS)中, 单个标记分子在焦体积内外的扩散会引起荧光随时间的波动, 通过将荧光涨落曲线拟合到适当的扩散模型, 可计算粒子数及其扩散系数; (C) 选择感兴趣区域(ROI), 用强激光束漂白, 并测量ROI中的荧光恢复时间; (D) 超分辨显微成像技术的原理示意图。FCS、SIM、STED、PALM和STORM同表1。

Figure 2 Technology of plasma membrane protein dynamics (A) From refraction to total internal reflection (TIR): the incident beam approaching the interface between two media with different indices of refraction n1 and n2 at an angle α will be refracted according to Snell’s law, beam propagation after the interface occurs thus at a different angle, if the incident angle α1 is greater than or equal to the critical angle α2, the beam does not propagate in the optically less-dense medium, but instead undergoes total reflection at the interface; (B) The diffusion of single labeled molecules in and out of the focal volume in fluorescence correlation spectroscopy (FCS) induces fluctuations in fluorescence with time, the number of particles and their diffusion coefficients can be calculated by fitting the fluorescence fluctuation curve to an adequate diffusion model; (C) A region of interest (ROI) is selected bleached with an intense laser beam, and the fluorescence recovery in the ROI is measured over time; (D) Schematic diagram of the principles of super-resolution microscopic imaging techniques. FCS, SIM, STED, PALM and STORM see Table 1.