Department of Biology , University of Osnabruck, Barbarastr. 11, 49076 Osnabrück, Germany.
Biol Chem. 2013 Sep;394(9):1097-113. doi: 10.1515/hsz-2012-0324.
Unraveling the spatio-temporal organization of dynamic cellular microcompartments requires live cell imaging techniques capable of resolving submicroscopic structures. While the resolution of traditional far-field fluorescence imaging techniques is limited by the diffraction barrier, several fluorescence-based microscopy techniques providing sub-100 nm resolution have become available during the past decade. Here, we briefly introduce the optical principles of these techniques and compare their capabilities and limitations with respect to spatial and temporal resolution as well as live cell capabilities. Moreover, we summarize how these techniques contributed to a better understanding of plasma membrane microdomains, the dynamic nanoscale organization of neuronal synapses and the sub-compartmentation of microorganisms. Based on these applications, we highlight complementarity of these techniques and their potential to address specific challenges in the context of dynamic cellular microcompartments, as well as the perspectives to overcome current limitations of these methods.
解析动态细胞微区室的时空组织需要能够解析亚微观结构的活细胞成像技术。虽然传统的远场荧光成像技术的分辨率受到衍射障碍的限制，但在过去十年中，已经出现了几种提供亚 100nm 分辨率的基于荧光的显微镜技术。在这里，我们简要介绍了这些技术的光学原理，并比较了它们在空间和时间分辨率以及活细胞能力方面的能力和局限性。此外，我们总结了这些技术如何有助于更好地理解质膜微区室、神经元突触的动态纳米尺度组织以及微生物的亚区室化。基于这些应用，我们强调了这些技术的互补性及其在动态细胞微区室背景下解决特定挑战的潜力，以及克服这些方法当前局限性的观点。
