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Microsc Res Tech. 2011 Jul;74(7):688-98. doi: 10.1002/jemt.20962. Epub 2010 Dec 03.

Single molecule microscopy methods for the study of DNA origami structures.

Microscopy research and technique

Victoria Birkedal, Mingdong Dong, Monika M Golas, Bjoern Sander, Ebbe Sloth Andersen, Kurt Vesterager Gothelf, Flemming Besenbacher, Jørgen Kjems

Affiliations

  1. iNANO Center, Aarhus University, 8000 Århus C, Denmark. [email protected]

PMID: 21698717 DOI: 10.1002/jemt.20962

Abstract

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.

Copyright © 2010 Wiley-Liss, Inc.

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