Fast and multiplexed superresolution imaging with DNA-PAINT-ERS
Fast and multiplexed superresolution imaging with DNA-PAINT-ERS
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DOI：
10.1038/s41467-020-18181-6
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发表时间：
2020-08-28
影响因子：
16.6
通讯作者：
Nan, Xiaolin
中科院分区：
文献类型：
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作者：
Civitci, Fehmi;Shangguan, Julia;Nan, Xiaolin
DNA点积累纳米级形貌成像（DNA-PAINT）有利于多路复用在超分辨率显微镜，但实际上是由缓慢的成像速度限制。为了解决这个问题，我们建议添加碳酸亚乙酯（EC）的成像缓冲液，序列重复的对接链，和对接链和亲和剂之间的间隔。统称为DNA-PAINT-ERS（E = EC，R =重复序列，S =间隔区），这些策略可以很容易地集成到当前的DNA-PAINT工作流程中，通过优化DNA杂交动力学和效率来加快成像速度和提高图像质量。我们证明了DNA-PAINT-ERS的快速，多路复用的超分辨率成像的一般适用性，使用以前验证的寡核苷酸结构进行轻微的修改。
DNA points accumulation for imaging in nanoscale topography (DNA-PAINT) facilitates multiplexing in superresolution microscopy but is practically limited by slow imaging speed. To address this issue, we propose the additions of ethylene carbonate (EC) to the imaging buffer, sequence repeats to the docking strand, and a spacer between the docking strand and the affinity agent. Collectively termed DNA-PAINT-ERS (E = EC, R = Repeating sequence, and S = Spacer), these strategies can be easily integrated into current DNA-PAINT workflows for both accelerated imaging speed and improved image quality through optimized DNA hybridization kinetics and efficiency. We demonstrate the general applicability of DNA-PAINT-ERS for fast, multiplexed superresolution imaging using previously validated oligonucleotide constructs with slight modifications.