Site-Specific Labeling of Affimers for DNA-PAINT Microscopy
Site-Specific Labeling of Affimers for DNA-PAINT Microscopy
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DOI：
10.1002/anie.201804020
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发表时间：
2018-08-20
影响因子：
16.6
通讯作者：
Jungmann, Ralf
中科院分区：
文献类型：
--
作者：
Schlichthaerle, Thomas;Eklund, Alexandra S.;Jungmann, Ralf
光学超分辨率技术允许低于光的经典衍射极限的荧光成像。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，这一显著成就并不容易转化为细胞成分的成像，因为目前的标记方法受到大标记尺寸（抗体）或小而有效的结合剂（纳米抗体、适体、遗传编码标签）的稀疏可用性的限制。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。我们使用肌动蛋白亲和聚体测定了我们的方法。小的DNA缀合的亲和结合剂可以为未来高效的多靶点超分辨率成像提供解决方案。
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.