Three-dimensional super-resolution imaging of the midplane protein FtsZ in live Caulobacter crescentus cells using astigmatism.

Three-dimensional super-resolution imaging of the midplane protein FtsZ in live Caulobacter crescentus cells using astigmatism.
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DOI:
10.1002/cphc.201100686
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发表时间:
2012-03
期刊:
Chemphyschem : a European journal of chemical physics and physical chemistry
影响因子:
--
通讯作者:
Moerner WE
Moerner WE
中科院分区:
其他
文献类型:
--
作者:
Biteen JS;Goley ED;Shapiro L;Moerner WE

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单分子超分辨率成像为纳米级成像提供了一种非侵入性方法,非常适合活细胞内准静态结构的研究。在这里,我们扩展的能力,在细菌细胞内的亚细胞特征的图像到三维的基础上,在成像路径中引入圆柱形透镜。我们研究了新月柄杆菌中的中平面蛋白FtsZ与超分辨率成像的基础上荧光蛋白的光开关和自然的聚合/解聚动力学FtsZ与Z-环。我们量化这些动力学,并确定FtsZ解聚时间为<100毫秒。在细胞周期的不同阶段的crescentus细胞,发现FtsZ超结构是动态的细胞周期,形成一个开放的形状在柄阶段和密集的焦点在分裂前阶段。
Single-molecule super-resolution imaging provides a non-invasive method for nanometer-scale imaging and is ideally suited to investigations of quasi-static structures within live cells. Here, we extend the ability to image subcellular features within bacteria cells to three dimensions based on the introduction of a cylindrical lens in the imaging pathway. We investigate the midplane protein FtsZ in Caulobacter crescentus with super-resolution imaging based on fluorescent-protein photo-switching and the natural polymerization/depolymerization dynamics of FtsZ associated with the Z-ring. We quantify these dynamics and determine the FtsZ depolymerization time to be <100 ms. We image the Z-ring in live and fixed C. crescentus cells at different stages of the cell cycle and find that the FtsZ superstructure is dynamic with the cell cycle, forming an open shape during the stalked stage and a dense focus during the pre-divisional stage.
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