Opportunities to Explore Plant Membrane Organization with Super-Resolution Microscopy
Opportunities to Explore Plant Membrane Organization with Super-Resolution Microscopy
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DOI：
10.1104/pp.110.161703
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发表时间：
2010-10-01
期刊：
影响因子：
7.4
通讯作者：
Ehrhardt, David W.
中科院分区：
文献类型：
--
作者：
Gutierrez, Ryan;Grossmann, Guido;Ehrhardt, David W.
电子显微镜和光学显微镜都是研究分子分布和细胞结构的重要工具。虽然与光学显微镜相比，电子显微镜具有更高的分辨率，但它容易因样品制备而产生伪影，并且只产生静态图像，这使得动态过程的分析具有挑战性。此外，在电子显微镜中进行特定分子标记和检测的方法也面临着显著的局限性。另一方面，光学显微镜擅长于特定的标记，特别是遗传编码标签，并且可以在分子和细胞器的动态可以可视化的活细胞中使用。然而，与电子显微镜相比，光学显微镜在检测单个蛋白质和蛋白质复合物以及在光衍射极限以下解析结构和分子分布的能力方面受到限制。最近的技术发展解决了电子显微镜和光显微镜的缺点，同时保留了它们的优点。引入遗传编码标签的新策略为电子显微镜带来了更大的分子特异性（回顾，见Giepmans， 2008），光探测器和成像方法的进步使得体内单分子检测成为可能，并提高了光学显微镜在衍射极限以下的分辨能力（图1）。在这里，我们讨论了光学显微镜的最新进展，以及它们如何应用于进一步我们对植物细胞结构和功能的理解，重点是质膜（PM）的横向组织。
Electron microscopy and light microscopy both have been essential tools for investigating molecular distribution and cell structure. While electron microscopy is capable of much higher resolution compared to light microscopy, it is prone to artifacts introduced by sample preparation and it produces only static images, making the analysis of dynamic processes challenging. In addition, methods for specific molecular labeling and detection in electron microscopy have faced significant limitations. Light microscopy, on the other hand, excels at specific labeling, particularly with genetically encoded tags, and can be used in living cells where the dynamics of molecules and organelles can be visualized. However, light microscopy has been limited in comparison to electron microscopy in its ability to detect single proteins and protein complexes and to resolve structure and molecular distributions below the diffraction limit of light. Recent technical developments address the shortcomings in both electron and light microscopy, while retaining their strengths. New strategies for introducing genetically encoded tags are bringing greater molecular specificity to electron microscopy (for review, see Giepmans, 2008), and advances in light detectors and imaging methods have permitted single molecule detection in vivo and have increased the resolving power of light microscopy below the diffraction limit (Fig. 1). Here, we discuss recent advances in light microscopy and how they can be applied to further our understanding of plant cell structure and function, with an emphasis on the lateral organization of the plasma membrane (PM).