Institute of Biochemistry, ETH Zurich, Switzerland.
Nat Methods. 2012 Jun;9(6):582-4. doi: 10.1038/nmeth.1991. Epub 2012 Apr 29.
We developed a method to use any GFP-tagged construct in single-molecule super-resolution microscopy. By targeting GFP with small, high-affinity antibodies coupled to organic dyes, we achieved nanometer spatial resolution and minimal linkage error when analyzing microtubules, living neurons and yeast cells. We show that in combination with libraries encoding GFP-tagged proteins, virtually any known protein can immediately be used in super-resolution microscopy and that simplified labeling schemes allow high-throughput super-resolution imaging.
我们开发了一种在单分子超分辨率显微镜中使用任何 GFP 标记构建体的方法。通过将 GFP 与偶联有机染料的小而高亲和力的抗体靶向,我们在分析微管、活神经元和酵母细胞时实现了纳米级空间分辨率和最小的连接错误。我们表明,与编码 GFP 标记蛋白的文库结合使用,实际上任何已知的蛋白都可以立即在超分辨率显微镜中使用,并且简化的标记方案允许高通量超分辨率成像。