Optically monitoring the mechanical assembly of single molecules
Optically monitoring the mechanical assembly of single molecules
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DOI：
10.1038/nnano.2008.333
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发表时间：
2009-01-01
影响因子：
38.3
通讯作者：
Gaub, Hermann E.
中科院分区：
文献类型：
--
作者：
Kufer, Stefan K.;Strackharn, Mathias;Gaub, Hermann E.
在单个分子水平上进行自下而上的组装需要最高的空间精度和有效的监测相结合。我们之前已经展示了如何‘剪切和粘贴’单分子‘，其他小组已经证明了有可能在光学显微镜下打破衍射极限(2-4)。在这里，我们展示了单分子剪切和粘贴表面组装(1)、全内反射荧光显微镜和原子力显微镜(5-8)的组合可以用来以明确的纳米级图案沉积单个荧光团，并且还可以以纳米级的精度实时监测这一过程。尽管图案的大小远低于显微镜的光学分辨率，但通过定位质心和检测荧光团的光漂白来识别单独的染料。通过这种方法的组合，单独的染料或标记的生物分子可以随意排列以实现特定的功能，例如耦合的荧光团系统或定制的酶级联，并以纳米级的精度进行监测。
Bottom-up assembly at the level of individual molecules requires a combination of utmost spatial precision and efficient monitoring. We have previously shown how to 'cut-and-paste' single molecules', and other groups have demonstrated that it is possible to beat the diffraction limit in optical microscopy(2-4). Here we show that a combination of single-molecule cut-and-paste surface assembly(1), total internal reflection fluorescence microscopy and atomic force microscopy(5-8) can be used to deposit individual fluorophores in well-defined nanoscale patterns and also to monitor the process in real time with nanometre precision. Although the size of the pattern is well below the optical resolution of the microscope, the individual dyes are identified by localizing the centroids and detecting the photobleaching of the fluorophores. With this combination of methods, individual dyes or labelled biomolecules can be arranged at will for specific functions, such as coupled fluorophore systems or tailored enzyme cascades, and monitored with nanoscale precision.