iNANO Center, Aarhus University, 8000 Århus C, Denmark.
Microsc Res Tech. 2011 Jul;74(7):688-98. doi: 10.1002/jemt.20962. Epub 2010 Dec 3.
Single molecule microscopy techniques play an important role in the investigation of advanced DNA structures such as those created by the DNA origami method. Three single molecule microscopy techniques are particularly interesting for the investigation of complex self-assembled three-dimensional (3D) DNA nanostructures, namely single molecule fluorescence microscopy, atomic force microscopy (AFM), and cryogenic transmission electron microscopy (cryo-EM). Here we discuss the strengths of these three techniques and demonstrate how their interplay can yield very important and unique new insights into the structure and conformation of advanced biological nanostructures. The applications of the three single molecule microscopy techniques are illustrated by focusing on a self-assembled DNA origami 3D box nanostructure. Its size and structure were studied by AFM and cryo-EM, while the lid opening, which can be controlled by the addition of oligonucleotide keys, was recorded by Förster/fluorescence resonance energy transfer (FRET) spectroscopy.
单分子显微镜技术在研究高级 DNA 结构（如 DNA 折纸方法所创建的结构）方面发挥着重要作用。三种单分子显微镜技术特别适用于研究复杂的自组装三维（3D）DNA 纳米结构，即单分子荧光显微镜、原子力显微镜（AFM）和低温透射电子显微镜（cryo-EM）。在这里，我们讨论了这三种技术的优势，并展示了它们的相互作用如何为高级生物纳米结构的结构和构象提供非常重要和独特的新见解。通过聚焦于自组装的 DNA 折纸 3D 盒纳米结构，说明了这三种单分子显微镜技术的应用。通过 AFM 和 cryo-EM 研究了其大小和结构，而通过Förster/荧光共振能量转移（FRET）光谱学记录了可以通过添加寡核苷酸键来控制的盖子打开。
