Super-resolution mbPAINT for optical localization of single-stranded DNA.

Super-resolution mbPAINT for optical localization of single-stranded DNA.
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DOI:
10.1021/am403984k
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发表时间:
2013-10-09
影响因子:
9.5
通讯作者:
Landes, Christy F.
Landes, Christy F.
中科院分区:
材料科学2区
文献类型:
--
作者:
Chen, Jixin;Bremauntz, Alberto;Kisley, Lydia;Shuang, Bo;Landes, Christy F.

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我们展示了应用超定位显微镜来识别单链DNA(SsDNA)的序列特定部分,序列分辨率为50个核苷酸,对应的空间分辨率为30 nm。超分辨率成像是使用一种单分子定位方法的变体实现的,该方法被称为用于纳米级地形成像的运动模糊点累积(MBPAINT)。目标单链DNA分子被固定在底物上。短的、染料标记的和互补的单链DNA分子随机结合到目标单链DNA上,重复的结合事件允许超分辨。序列特异性通过使用对照的、非互补的探针来证明。这些结果支持了使用相对便宜的短单链DNA来鉴定基因序列特异性的可能性,与现有方法相比,分辨率有所提高。
We demonstrate the application of super-localization microscopy to identify sequence-specific portions of single-stranded DNA (ssDNA) with sequence resolution of 50 nucleotides, corresponding to a spatial resolution of 30 nm. Super-resolution imaging was achieved using a variation of a single-molecule localization method, termed as ‘motion blur’ point accumulation for imaging in nanoscale topography (mbPAINT). The target ssDNA molecules were immobilized on the substrate. Short, dye-labeled, and complementary ssDNA molecules stochastically bound to the target ssDNA, with repeated binding events allowing super-resolution. Sequence specificity was demonstrated via the use of a control, non-complementary probe. The results support the possibility of employing relatively inexpensive short ssDNAs to identify gene sequence specificity with improved resolution in comparison to the existing methods.
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