title: Multicolor super-resolution imaging with photo-switchable fluorescent probes.
abstract: Recent advances in far-field optical nanoscopy have enabled fluorescence imaging with a spatial resolution of 20 to 50 nanometers. Multicolor super-resolution imaging, however, remains a challenging task. Here, we introduce a family of photo-switchable fluorescent probes and demonstrate multicolor stochastic optical reconstruction microscopy (STORM). Each probe consists of a photo-switchable "reporter" fluorophore that can be cycled between fluorescent and dark states, and an "activator" that facilitates photo-activation of the reporter. Combinatorial pairing of reporters and activators allows the creation of probes with many distinct colors. Iterative, color-specific activation of sparse subsets of these probes allows their localization with nanometer accuracy, enabling the construction of a super-resolution STORM image. Using this approach, we demonstrate multicolor imaging of DNA model samples and mammalian cells with 20- to 30-nanometer resolution. This technique will facilitate direct visualization of molecular interactions at the nanometer scale.
authors: School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA.
authors: Mark
authors: M
authors: Bates
authors: Bo
authors: B
authors: Huang
authors: Graham T
authors: GT
authors: Dempsey
authors: Xiaowei
authors: X
authors: Zhuang
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D022163:Clathrin-Coated Vesicles; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D005456:Fluorescent Dyes; D008856:Microscopy, Fluorescence / Q000379:methods; D008870:Microtubules; D036103:Nanotechnology
chemical_list: D015342:DNA Probes; D005456:Fluorescent Dyes; D004247:DNA
references: Proc Natl Acad Sci U S A. 1978 Apr;75(4):1820-4
references: 417343
references: Biophys J. 2002 May;82(5):2775-83
references: 11964263
references: Proc Natl Acad Sci U S A. 2006 Jul 18;103(29):10929-34
references: 16829583
references: Science. 2005 Jun 3;308(5727):1469-72
references: 15817813
references: J Cell Biol. 1989 Feb;108(2):389-400
references: 2563728
references: Science. 2006 Apr 14;312(5771):217-24
references: 16614209
references: J Phys Chem B. 2005 Dec 29;109(51):24220-4
references: 16375416
references: Science. 2004 Sep 17;305(5691):1782-6
references: 15375270
references: Nat Biotechnol. 2003 Nov;21(11):1347-55
references: 14595362
references: Nature. 2005 Nov 24;438(7067):460-5
references: 16284617
references: Proc Natl Acad Sci U S A. 1967 Aug;58(2):719-26
references: 5233469
references: Biophys J. 2004 Aug;87(2):1328-37
references: 15298935
references: Curr Opin Biotechnol. 2005 Feb;16(1):35-40
references: 15722013
references: Science. 1999 Mar 12;283(5408):1670-6
references: 10073924
references: Phys Rev Lett. 2005 Mar 18;94(10):108101
references: 15783528
references: Proc Natl Acad Sci U S A. 2005 Sep 13;102(37):13081-6
references: 16141335
references: Proc Natl Acad Sci U S A. 2004 Apr 27;101(17):6462-5
references: 15096603
references: Biophys J. 2006 Dec 1;91(11):4258-72
references: 16980368
references: Science. 2007 May 25;316(5828):1153-8
references: 17525330
references: Opt Express. 2005 Sep 5;13(18):7052-62
references: 19498727
references: Nature. 2006 Jan 5;439(7072):105-8
references: 16397502
references: Biophys J. 2007 Apr 15;92(8):L67-9
references: 17307826
references: Science. 2003 Jun 27;300(5628):2061-5
references: 12791999
references: Proc Natl Acad Sci U S A. 2006 Dec 12;103(50):18911-6
references: 17142314
references: Science. 2006 Sep 15;313(5793):1642-5
references: 16902090
references: J Am Chem Soc. 2005 Mar 23;127(11):3801-6
references: 15771514
references: Proc Natl Acad Sci U S A. 2004 Aug 3;101(31):11298-303
references: 15277661
references: Nat Methods. 2006 Oct;3(10):793-5
references: 16896339