Photobleaching Enables Super-resolution Imaging of the FtsZ Ring in the Cyanobacterium Prochlorococcus

被引:0
|
作者
Zhan, Yuanchao [1 ]
Liu, Yaxin [1 ]
Zeng, Qinglu [1 ,2 ,3 ]
机构
[1] Hong Kong Univ Sci & Technol, Dept Ocean Sci, Hong Kong, Peoples R China
[2] Hong Kong Univ Sci & Technol, Div Life Sci, Hong Kong, Peoples R China
[3] HKUST Shenzhen Res Inst, Shenzhen, Peoples R China
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2018年 / 141期
基金
中国国家自然科学基金;
关键词
Immunology and Infection; Issue; 141; STORM; photobleaching; cyanobacterium; Prochlorococcus; super-resolution imaging; three-dimensional; FtsZ ring; cell division; OPTICAL RECONSTRUCTION MICROSCOPY;
D O I
10.3791/58603
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Super-resolution microscopy has been widely used to study protein interactions and subcellular structures in many organisms. In photosynthetic organisms, however, the lateral resolution of super-resolution imaging is only similar to 100 nm. The low resolution is mainly due to the high autofluorescence background of photosynthetic cells caused by high-intensity lasers that are required for super-resolution imaging, such as stochastic optical reconstruction microscopy (STORM). Here, we describe a photobleaching-assisted STORM method which was developed recently for imaging the marine picocyanobacterium Prochlorococcus. After photobleaching, the autofluorescence of Prochlorococcus is effectively reduced so that STORM can be performed with a lateral resolution of similar to 10 nm. Using this method, we acquire the in vivo three-dimensional (3-D) organization of the FtsZ protein and characterize four different FtsZ ring morphologies during the cell cycle of Prochlorococcus. The method we describe here might be adopted for the super-resolution imaging of other photosynthetic organisms.
引用
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页数:10
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