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	Highly photostable, reversibly photoswitchable fluorescent protein with high contrast ratio for live-cell superresolution microscopy
	Zhang, Xi1,2; Zhang, Mingshu1,3; Li, Dong4,5; He, Wenting1; Peng, Jianxin2; Betzig, Eric4; Xu, Pingyong1,3,6
刊名	Proceedings of the national academy of sciences of the united states of america
	2016-09-13
卷号	113期号:37页码:10364-10369
关键词	Fluorescent protein Superresolution Microscopy Live-cell imaging Skylan-ns
ISSN号	0027-8424
DOI	10.1073/pnas.1611038113
通讯作者	Betzig, eric(betzige@janelia.hhmi.org) ; Xu, pingyong(pyxu@ibp.ac.cn)
英文摘要	Two long-standing problems for superresolution (sr) fluorescence microscopy are high illumination intensity and long acquisition time, which significantly hamper its application for live-cell imaging. reversibly photoswitchable fluorescent proteins (rsfps) have made it possible to dramatically lower the illumination intensities in saturated depletion-based sr techniques, such as saturated depletion nonlinear structured illumination microscopy (nl-sim) and reversible saturable optical fluorescence transition microscopy. the characteristics of rsfps most critical for sr live-cell imaging include, first, the integrated fluorescence signal across each switching cycle, which depends upon the absorption cross-section, effective quantum yield, and characteristic switching time from the fluorescent "on" to "off" state; second, the fluorescence contrast ratio of on/off states; and third, the photostability under excitation and depletion. up to now, the rsfps of the dronpa and rsegfp (reversibly switchable egfp) families have been exploited for sr imaging. however, their limited number of switching cycles, relatively low fluorescence signal, and poor contrast ratio under physiological conditions ultimately restrict their utility in time-lapse live-cell imaging and their ability to reach the desired resolution at a reasonable signal-to-noise ratio. here, we present a truly monomeric rsfp, skylan-ns, whose properties are optimized for the recently developed patterned activation nl-sim, which enables low-intensity (similar to 100 w/cm(2)) live-cell sr imaging at similar to 60-nm resolution at subsecond acquisition times for tens of time points over broad field of view.
WOS关键词	STRUCTURED-ILLUMINATION MICROSCOPY ; PATTERNED EXCITATION MICROSCOPY ; LOCALIZATION MICROSCOPY ; RESOLUTION LIMIT ; NANOSCOPY ; BREAKING ; BRIGHT ; DRONPA
WOS研究方向	Science & Technology - Other Topics
WOS类目	Multidisciplinary Sciences
语种	英语
出版者	NATL ACAD SCIENCES
WOS记录号	WOS:000383092000048
内容类型	期刊论文
URI标识	http://www.corc.org.cn/handle/1471x/2376269
专题	中国科学院大学
通讯作者	Betzig, Eric; Xu, Pingyong
作者单位	1.Chinese Acad Sci, Inst Biophys, Key Lab RNA Biol, Beijing 100101, Peoples R China 2.Cent China Normal Univ, Sch Life Sci, Inst Entomol, Wuhan 430079, Hubei, Peoples R China 3.Chinese Acad Sci, Beijing Key Lab Noncoding RNA, Inst Biophys, Beijing 100101, Peoples R China 4.Howard Hughes Med Inst, Janelia Res Campus, Ashburn, VA 20147 USA 5.Chinese Acad Sci, Inst Biophys, CAS Ctr Excellence Biomacromol, Natl Lab Biomacromol, Beijing 100101, Peoples R China 6.Univ Chinese Acad Sci, Coll Life Sci, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Zhang, Xi,Zhang, Mingshu,Li, Dong,et al. Highly photostable, reversibly photoswitchable fluorescent protein with high contrast ratio for live-cell superresolution microscopy[J]. Proceedings of the national academy of sciences of the united states of america,2016,113(37):10364-10369.
APA	Zhang, Xi.,Zhang, Mingshu.,Li, Dong.,He, Wenting.,Peng, Jianxin.,...&Xu, Pingyong.(2016).Highly photostable, reversibly photoswitchable fluorescent protein with high contrast ratio for live-cell superresolution microscopy.Proceedings of the national academy of sciences of the united states of america,113(37),10364-10369.
MLA	Zhang, Xi,et al."Highly photostable, reversibly photoswitchable fluorescent protein with high contrast ratio for live-cell superresolution microscopy".Proceedings of the national academy of sciences of the united states of america 113.37(2016):10364-10369.

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