Development of a Reversibly Switchable Fluorescent Protein for   Super-Resolution Optical Fluctuation Imaging (SOFI)

doi:10.1021/nn5064387

CORC > 北京大学 > 生命科学学院

	Development of a Reversibly Switchable Fluorescent Protein for Super-Resolution Optical Fluctuation Imaging (SOFI)
	Zhang, Xi ; Chen, Xuanze ; Zeng, Zhiping ; Zhang, Mingshu ; Sun, Yujie ; Xi, Peng ; Peng, Jianxin ; Xu, Pingyong
刊名	acs nano
	2015
关键词	reversibly switchable fluorescent protein super-resolution microscopy Skylan-S protein engineering live-cell imaging SOFI MICROSCOPY RESOLUTION NANOSCOPY STORM LIMIT GFP
DOI	10.1021/nn5064387
英文摘要	Reversibly switchable fluorescent proteins (RSFPs) can be effectively used for super-resolution optical fluctuation imaging (SOFI) based on the switching and fluctuation of single molecules. Several properties of RSFPs strongly influence the quality of SOFI images. These properties include (i) the averaged fluorescence intensity in the fluctuation state, (ii) the on/off contrast ratio, (iii) the photostability, and (iv) the oligomerization tendency. The first three properties determine the fluctuation range of the imaged pixels and the SOFI signal, which are of essential importance to the spatial resolution, and the last may lead to artificial aggregation of target proteins. The RSFPs that are currently used for SOFI are low in averaged fluorescence intensity in the fluctuation state, photostability, and on/off contrast ratio, thereby limiting the range of application of SOFI in biological super-resolution imaging. In this study, we developed a novel monomeric green RSFP termed Skylan-S, which features very high photostability, contrast ratio, and averaged fluorescence intensity in the fluctuation state. Taking advantage of the excellent optical properties of Skylan-S, a 4-fold improvement in the fluctuation range of the imaged pixels and higher SOFI resolution can be obtained compared with Dronpa. Furthermore, super-resolution imaging of the actin or tubulin structures and clathrin-coated pits (CCPs) in living U2OS cells labeled with Skylan-S was demonstrated using the SOFI technique. Overall, Skylan-S developed with outstanding photochemical properties is promising for long-time SOH imaging with high spatial-temporal resolution.; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000351791800041&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; Chemistry, Multidisciplinary; Chemistry, Physical; Nanoscience & Nanotechnology; Materials Science, Multidisciplinary; SCI(E); EI; PubMed; 11; ARTICLE; xipeng@pku.edu.cn; jxpeng@mail.ccnu.edu.cn; pyxu@ibp.ac.cn; 3; 2659-2667; 9
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/188282]
专题	生命科学学院工学院
推荐引用方式 GB/T 7714	Zhang, Xi,Chen, Xuanze,Zeng, Zhiping,et al. Development of a Reversibly Switchable Fluorescent Protein for Super-Resolution Optical Fluctuation Imaging (SOFI)[J]. acs nano,2015.
APA	Zhang, Xi.,Chen, Xuanze.,Zeng, Zhiping.,Zhang, Mingshu.,Sun, Yujie.,...&Xu, Pingyong.(2015).Development of a Reversibly Switchable Fluorescent Protein for Super-Resolution Optical Fluctuation Imaging (SOFI).acs nano.
MLA	Zhang, Xi,et al."Development of a Reversibly Switchable Fluorescent Protein for Super-Resolution Optical Fluctuation Imaging (SOFI)".acs nano (2015).