Visualization of barriers and obstacles to molecular diffusion in live cells by spatial pair-cross-correlation in two dimensions
Visualization of barriers and obstacles to molecular diffusion in live cells by spatial pair-cross-correlation in two dimensions
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DOI：
10.1364/boe.9.000303
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发表时间：
2018-01-01
影响因子：
3.4
通讯作者：
Gratton, Enrico
中科院分区：
文献类型：
--
作者：
Malacrida, Leonel;Hedde, Per Niklas;Gratton, Enrico
尽管最近在光学超分辨率方面取得了进展，但我们缺乏一种方法，可以可视化细胞质或细胞核中扩散分子的路径。荧光相关光谱（FCS）通过在单个点处对许多分子随时间的行为进行平均来提供单分子水平的分子动力学，从而实现非常好的统计，但仅在细胞中的一个点处。早期的基于图像的方法，包括光栅扫描和时空图像相关需要在相对较大的区域上进行空间平均，从而损害空间分辨率。在这里，我们使用二维空间对互相关（2D-pCF）来获得相对高分辨率的活细胞中分子扩散动力学和运输图像。2D-pCF方法通过图像中特定点处的强度波动的互相关来测量颗粒从一个位置到另一个位置的时间。因此，创建了分子遵循的平均路径的视觉图。(c)根据OSA开放获取出版协议的条款，2017年美国光学学会
Despite recent advances in optical super-resolution, we lack a method that can visualize the path followed by diffusing molecules in the cytoplasm or in the nucleus of cells. Fluorescence correlation spectroscopy (FCS) provides molecular dynamics at the single molecule level by averaging the behavior of many molecules over time at a single spot, thus achieving very good statistics but at only one point in the cell. Earlier image-based methods including raster-scan and spatiotemporal image correlation need spatial averaging over relatively large areas, thus compromising spatial resolution. Here, we use spatial pair-cross-correlation in two dimensions (2D-pCF) to obtain relatively high resolution images of molecular diffusion dynamics and transport in live cells. The 2D-pCF method measures the time for a particle to go from one location to another by cross-correlating the intensity fluctuations at, specific points in an image. Hence, a visual map of the average path followed by molecules is created. (c) 2017 Optical Society of America under the terms of the OSA Open Access Publishing Agreement