Realizing the Full Potential of Advanced Microscopy Approaches for Interrogating Plant-Microbe Interactions
Realizing the Full Potential of Advanced Microscopy Approaches for Interrogating Plant-Microbe Interactions
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DOI：
10.1094/mpmi-10-22-0208-fi
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发表时间：
2023-04-01
影响因子：
3.5
通讯作者：
Berg，Howard
中科院分区：
文献类型：
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作者：
Czymmek，Kirk J.;Duncan，Keith E.;Berg，Howard
几个世纪以来，显微镜一直是洞察和发现植物-微生物相互作用的基本工具。从经典的光学显微镜和电子显微镜到相应的样品制备和细胞对比剂的专门方法，这些方法已经成为植物和微生物学科学家可视化、探测和了解宿主-微生物关系本质的工具包中的常规组件。在过去的三十年里，随着电子断层扫描技术的发展，尤其是共聚焦技术的发展，三维透视技术继续提供组织和细胞现象的显著清晰度和空间细节。共焦显微镜和电子显微镜提供了如今在中型和大型机构中司空见惯的新发现。然而，许多其他尖端技术和样品制备工作流程相对来说还没有被开发，但在我们对致病、有益和共生的植物-微生物相互作用的内部工作原理的理解方面提供了前所未有的巨大潜力。在这里，我们重点介绍当代先进的植物-微生物系统成像平台的主要应用、优势和挑战，特别是最近开发的几种方法，如薄片、单分子、超分辨率和自适应光学显微镜，以及环境和低温体积电子显微镜、X射线显微镜和低温电子断层扫描。此外，还将审查补充样品制备方法的潜力，如光学清除、膨胀显微镜和多重成像。我们的最终目标是激发人们对这些强大的尖端技术的认识，并促进它们的适当应用和采用，以解决该领域尚未解决的重要生物学问题。版权所有©2023作者(S)。这是一篇在CC by 4.0国际许可证下分发的开放获取文章。
Microscopy has served as a fundamental tool for insight and discovery in plant-microbe interactions for centuries. From classical light and electron microscopy to corresponding specialized methods for sample preparation and cellular contrasting agents, these approaches have become routine components in the toolkit of plant and microbiology scientists alike to visualize, probe and understand the nature of host-microbe relationships. Over the last three decades, three-dimensional perspectives led by the development of electron tomography, and especially, confocal techniques continue to provide remarkable clarity and spatial detail of tissue and cellular phenomena. Confocal and electron microscopy provide novel revelations that are now commonplace in medium and large institutions. However, many other cutting-edge technologies and sample preparation workflows are relatively unexploited yet offer tremendous potential for unprecedented advancement in our understanding of the inner workings of pathogenic, beneficial, and symbiotic plant-microbe interactions. Here, we highlight key applications, benefits, and challenges of contemporary advanced imaging platforms for plant-microbe systems with special emphasis on several recently developed approaches, such as light-sheet, single molecule, super-resolution, and adaptive optics microscopy, as well as ambient and cryo-volume electron microscopy, X-ray microscopy, and cryo-electron tomography. Furthermore, the potential for complementary sample preparation methodologies, such as optical clearing, expansion microscopy, and multiplex imaging, will be reviewed. Our ultimate goal is to stimulate awareness of these powerful cutting-edge technologies and facilitate their appropriate application and adoption to solve important and unresolved biological questions in the field.Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.