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Site‐Specific Labeling of Affimers for DNA‐PAINT Microscopy
Angewandte Chemie International Edition ( IF 16.6 ) Pub Date : 2018-07-15 , DOI: 10.1002/anie.201804020
Thomas Schlichthaerle 1, 2 , Alexandra S Eklund 1, 2 , Florian Schueder 1, 2 , Maximilian T Strauss 1, 2 , Christian Tiede 3, 4 , Alistair Curd 3, 4 , Jonas Ries 5 , Michelle Peckham 3, 4 , Darren C Tomlinson 3, 4 , Ralf Jungmann 1, 2
Affiliation  

Optical super‐resolution techniques allow fluorescence imaging below the classical diffraction limit of light. From a technology standpoint, recent methods are approaching molecular‐scale spatial resolution. However, this remarkable achievement is not easily translated to imaging of cellular components, since current labeling approaches are limited by either large label sizes (antibodies) or the sparse availability of small and efficient binders (nanobodies, aptamers, genetically‐encoded tags). In this work, we combined recently developed Affimer reagents with site‐specific DNA modification for high‐efficiency labeling and imaging using DNA‐PAINT. We assayed our approach using an actin Affimer. The small DNA‐conjugated affinity binders could provide a solution for efficient multitarget super‐resolution imaging in the future.

中文翻译:

用于 DNA-PAINT 显微镜的亲和物的位点特异性标记

光学超分辨率技术允许在光的经典衍射极限以下进行荧光成像。从技术的角度来看,最近的方法正在接近分子尺度的空间分辨率。然而,这一非凡的成就并不容易转化为细胞成分的成像,因为当前的标记方法受到大标签(抗体)或小而有效的结合剂(纳米抗体、适体、基因编码标签)的稀少可用性的限制。在这项工作中,我们将最近开发的 Affimer 试剂与位点特异性 DNA 修饰相结合,使用 DNA-PAINT 进行高效标记和成像。我们使用肌动蛋白 Affimer 分析了我们的方法。小 DNA 结合的亲和结合剂可以为未来的高效多目标超分辨率成像提供解决方案。
更新日期:2018-07-15
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