Department of Physics and Chemistry, and MEMPHYS - Center for Biomembrane Physics, University of Southern Denmark, Campusvej 55, DK-5230 Odense M, Denmark.
Curr Protein Pept Sci. 2011 Dec;12(8):699-713. doi: 10.2174/138920311798841672.
Single particle tracking (SPT) enables light microscopy at a sub-diffraction limited spatial resolution by a combination of imaging at low molecular labeling densities and computational image processing. SPT and related single molecule imaging techniques have found a rapidly expanded use within the life sciences. This expanded use is due to an increased demand and requisite for developing a comprehensive understanding of the spatial dynamics of bio-molecular interactions at a spatial scale that is equivalent to the size of the molecules themselves, as well as by the emergence of new imaging techniques and probes that have made historically very demanding and specialized bio-imaging techniques more easily accessible and achievable. SPT has in particular found extensive use for analyzing the molecular organization of biological membranes. From these and other studies using complementary techniques it has been determined that the organization of native plasma membranes is heterogeneous over a very large range of spatial and temporal scales. The observed heterogeneities in the organization have the practical consequence that the SPT results in investigations of native plasma membranes are time dependent. Furthermore, because the accessible time dynamics, and also the spatial resolution, in an SPT experiment is mainly dependent on the luminous brightness and photostability of the particular SPT probe that is used, available SPT results are ultimately dependent on the SPT probes. The focus of this review is on the impact that the SPT probe has on the experimental results in SPT.
单颗粒跟踪 (SPT) 通过在低分子标记密度下成像和计算图像处理的组合，实现了亚衍射极限空间分辨率的荧光显微镜。SPT 和相关的单分子成像技术在生命科学中得到了快速扩展的应用。这种扩展的应用是由于对发展对生物分子相互作用的空间动力学的全面理解的需求和必要性的增加，这种空间动力学的空间尺度相当于分子本身的大小，以及由于新的成像技术和探针的出现，这些技术使得历史上非常苛刻和专门的生物成像技术更容易获得和实现。SPT 特别用于分析生物膜的分子组织。从这些和其他使用互补技术的研究中可以确定，天然质膜的组织在非常大的空间和时间尺度上是不均匀的。观察到的组织不均匀性具有实际的后果，即对天然质膜进行 SPT 研究的结果是时间依赖性的。此外，由于在 SPT 实验中可访问的时间动力学以及空间分辨率主要取决于所使用的特定 SPT 探针的发光亮度和光稳定性，因此可用的 SPT 结果最终取决于 SPT 探针。本综述的重点是 SPT 探针对 SPT 实验结果的影响。
