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用于植物有丝分裂微管生物成像的先进显微镜方法。

Advanced microscopy methods for bioimaging of mitotic microtubules in plants.

作者信息

Vyplelová Petra, Ovečka Miroslav, Komis George, Šamaj Jozef

机构信息

Department of Cell Biology, Centre of the Region Haná for Biotechnological and Agricultural Research, Palacký University Olomouc, Olomouc, Czech Republic.

Department of Cell Biology, Centre of the Region Haná for Biotechnological and Agricultural Research, Palacký University Olomouc, Olomouc, Czech Republic.

出版信息

Methods Cell Biol. 2018;145:129-158. doi: 10.1016/bs.mcb.2018.03.019. Epub 2018 Apr 26.

Abstract

Mitotic cell division in plants is a dynamic process playing a key role in plant morphogenesis, growth, and development. Since progress of mitosis is highly sensitive to external stresses, documentation of mitotic cell division in living plants requires fast and gentle live-cell imaging microscopy methods and suitable sample preparation procedures. This chapter describes, both theoretically and practically, currently used advanced microscopy methods for the live-cell visualization of the entire process of plant mitosis. These methods include microscopy modalities based on spinning disk, Airyscan confocal laser scanning, structured illumination, and light-sheet bioimaging of tissues or whole plant organs with diverse spatiotemporal resolution. Examples are provided from studies of mitotic cell division using microtubule molecular markers in the model plant Arabidopsis thaliana, and from deep imaging of mitotic microtubules in robust plant samples, such as legume crop species Medicago sativa.

摘要

植物中的有丝分裂细胞分裂是一个动态过程,在植物形态发生、生长和发育中起着关键作用。由于有丝分裂的进程对外部压力高度敏感,记录活植物中的有丝分裂细胞分裂需要快速且温和的活细胞成像显微镜方法以及合适的样品制备程序。本章从理论和实践两方面描述了目前用于植物有丝分裂全过程活细胞可视化的先进显微镜方法。这些方法包括基于转盘、Airyscan共聚焦激光扫描、结构照明以及对组织或整个植物器官进行具有不同时空分辨率的光片生物成像的显微镜模式。文中提供了在模式植物拟南芥中使用微管分子标记研究有丝分裂细胞分裂的例子,以及在诸如豆科作物紫花苜蓿等健壮植物样品中有丝分裂微管的深度成像的例子。

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