Hinde Elizabeth, Cardarelli Francesco
Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine, CA, USA.
Center for Nanotechnology Innovation @NEST, Istituto Italiano di Tecnologia, Piazza San Silvestro 12, 56127, Pisa, Italy.
Biophys Rev. 2011 Sep;3(3):119. doi: 10.1007/s12551-011-0051-x. Epub 2011 Jul 19.
No methods proposed thus far have the capability to measure molecular flow in live cells at the single molecule level. Here, we review the potentiality of a newly established method based on the spatial correlation of fluorescence fluctuations at a pair of points in the sample (pair correlation method). The pair correlation function (pCF) offers a unique tool to probe the directionality of intracellular traffic, by measuring the accessibility of the cellular landscape and its role in determining the diffusive routes adopted by molecules. The sensitivity of the pCF method toward detection of barriers means that different structural elements of the cell can be tested in terms of penetrability and mechanisms of regulation imparted on molecular flow. This has been recently demonstrated in a series of studies looking at molecular transport inside live cells. Here, we will review the theory behind detection of barriers to molecular flow, the rules to interpret pCF data, and relevant applications to intracellular transport.
迄今为止提出的方法都无法在单分子水平上测量活细胞中的分子流动。在此，我们综述一种基于样品中一对点处荧光涨落的空间相关性的新方法（对相关方法）的潜力。对相关函数（pCF）提供了一种独特的工具，通过测量细胞环境的可达性及其在确定分子所采用的扩散途径中的作用，来探测细胞内运输的方向性。pCF方法对屏障检测的灵敏度意味着，可以根据细胞不同结构元件对分子流动的渗透性和调节机制进行测试。最近在一系列关于活细胞内分子运输的研究中已经证明了这一点。在此，我们将综述分子流动屏障检测背后的理论、解释pCF数据的规则以及细胞内运输的相关应用。
