Inferring diffusion in single live cells at the single-molecule level
Inferring diffusion in single live cells at the single-molecule level
复制标题
DOI：
10.1098/rstb.2012.0029
复制
发表时间：
2013-02-05
影响因子：
6.3
通讯作者：
Leake, Mark C.
中科院分区：
文献类型：
--
作者：
Robson, Alex;Burrage, Kevin;Leake, Mark C.
活细胞内分子的运动是生物过程的基本特征。在单分子和单细胞水平上观察和分析体内分子扩散细节的能力可以为理解扩散分子的分子结构和分子扩散的纳米级环境提供重要的见解。用于监测活细胞中的动态分子定位的工具的选择是荧光显微镜，特别是这样结合全内反射荧光与使用荧光蛋白（FP）的记者在提供特殊的成像对比度在相对生理条件下的细胞膜中的动态过程相比，竞争的单分子技术。存在几种不同的复杂扩散模式，由于潜在的随机行为，在分子水平上将这些模式彼此区分是具有挑战性的。传统上使用跟踪粒子的均方位移进行分析;然而，由于物理不稳定性，这通常需要比单个FP轨迹典型的更多的数据点。这里提出的是一种新的方法，允许强大的贝叶斯排名的扩散过程，以区分多个复杂的模式概率。这是一种计算方法，生物学家可以用来理解活细胞中的单分子特征。
The movement of molecules inside living cells is a fundamental feature of biological processes. The ability to both observe and analyse the details of molecular diffusion in vivo at the single-molecule and single-cell level can add significant insight into understanding molecular architectures of diffusing molecules and the nanoscale environment in which the molecules diffuse. The tool of choice for monitoring dynamic molecular localization in live cells is fluorescence microscopy, especially so combining total internal reflection fluorescence with the use of fluorescent protein (FP) reporters in offering exceptional imaging contrast for dynamic processes in the cell membrane under relatively physiological conditions compared with competing single-molecule techniques. There exist several different complex modes of diffusion, and discriminating these from each other is challenging at the molecular level owing to underlying stochastic behaviour. Analysis is traditionally performed using mean square displacements of tracked particles; however, this generally requires more data points than is typical for single FP tracks owing to photophysical instability. Presented here is a novel approach allowing robust Bayesian ranking of diffusion processes to discriminate multiple complex modes probabilistically. It is a computational approach that biologists can use to understand single-molecule features in live cells.