Department of Chemistry and ‡Department of Electrical and Computer Engineering, Rice University , Houston, Texas 77251-1892, United States .
ACS Appl Mater Interfaces. 2013 Oct 9;5(19):9338-43. doi: 10.1021/am403984k. Epub 2013 Sep 27.
We demonstrate the application of superlocalization 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, noncomplementary 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.
我们展示了超本地化显微镜在识别单链 DNA（ssDNA）序列特异性部分方面的应用，其序列分辨率为 50 个核苷酸，对应于 30nm 的空间分辨率。超分辨率成像使用一种单分子定位方法的变体来实现，该方法称为“用于纳米级形貌成像的运动模糊点积累”（mbPAINT）。目标 ssDNA 分子固定在基底上。短的、标记染料的、互补的 ssDNA 分子随机结合到目标 ssDNA 上，通过重复的结合事件实现超分辨率。通过使用对照非互补探针证明了序列特异性。结果支持了使用相对便宜的短 ssDNA 来识别基因序列特异性的可能性，与现有方法相比，分辨率得到了提高。
