Super-resolution fingerprinting detects chemical reactions and idiosyncrasies of single DNA pegboards.

Super-resolution fingerprinting detects chemical reactions and idiosyncrasies of single DNA pegboards.
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DOI:
10.1021/nl304415b
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发表时间:
2013-02-13
期刊:
影响因子:
10.8
通讯作者:
Walter NG
Walter NG
中科院分区:
材料科学1区
文献类型:
--
作者:
Johnson-Buck A;Nangreave J;Kim DN;Bathe M;Yan H;Walter NG

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我们采用单粒子荧光纳米显微镜技术PAINT(点积累的纳米级拓扑结构成像),使用特定位点的DNA探针,以获得二维密度图的特定功能图案的纳米级DNA折纸钉板。我们发现,PAINT的定位精度约为10 nm,足以可靠地区分密集(>104个特征μm−2)的亚100 nm寡核苷酸特征模式。我们采用双色PAINT来跟踪酶催化的对单个折纸的特征的修饰,并显示单个纳米钉板表现出稳定的、空间异质的探针结合模式或“指纹”。最后,我们提出的实验和建模证据表明,这些指纹可能会出现局部调制的探针结合动力学的特征间距的变化。我们的研究突出了荧光纳米显微镜的力量,对与溶液中试剂相互作用和定位的单个软纳米器件进行质量控制。
We employ the single-particle fluorescence nanoscopy technique PAINT (points accumulation for imaging in nanoscale topography) using site-specific DNA probes to acquire two-dimensional density maps of specific features patterned on nanoscale DNA origami pegboards. We show that PAINT has a localization accuracy of ~10 nm that is sufficient to reliably distinguish dense (>104 features μm−2) sub-100-nm patterns of oligonucleotide features. We employ two-color PAINT to follow enzyme-catalyzed modification of features on individual origami, and to show that single nano-pegboards exhibit stable, spatially heterogeneous probe-binding patterns, or “fingerprints.” Finally, we present experimental and modeling evidence suggesting that these fingerprints may arise from feature spacing variations that locally modulate the probe binding kinetics. Our study highlights the power of fluorescence nanoscopy to perform quality control on individual soft nanodevices that interact with and position reagents in solution.
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