Single Molecule Microscopy Methods for the Study of DNA Origami Structures

被引：15

作者：

Birkedal, Victoria ^{[1
]}

Dong, Mingdong ^{[1
,2
]}

Golas, Monika M. ^{[3
,4
]}

Sander, Bjoern ^{[4
,5
]}

Andersen, Ebbe Sloth ^{[1
,2
,6
]}

Gothelf, Kurt Vesterager ^{[1
,2
,7
]}

Besenbacher, Flemming ^{[1
,2
,8
]}

Kjems, Jorgen ^{[1
,2
,6
]}

机构：

[1] Aarhus Univ, iNANO Ctr, DK-8000 Aarhus C, Denmark

[2] Aarhus Univ, Ctr DNA Nanotechnol, DK-8000 Aarhus C, Denmark

[3] Aarhus Univ, Water & Salt Res Ctr, Inst Anat, DK-8000 Aarhus C, Denmark

[4] Aarhus Univ, Ctr Stochast Geometry & Adv Bioimaging, DK-8000 Aarhus C, Denmark

[5] Aarhus Univ, Inst Clin Med, Stereol & Electron Microscopy Lab, Inst Clin Med, DK-8000 Aarhus C, Denmark

[6] Aarhus Univ, Dept Mol Biol, DK-8000 Aarhus C, Denmark

[7] Aarhus Univ, Dept Chem, DK-8000 Aarhus C, Denmark

[8] Aarhus Univ, Dept Phys & Astron, DK-8000 Aarhus C, Denmark

来源：

MICROSCOPY RESEARCH AND TECHNIQUE | 2011年 / 74卷 / 07期

基金：

新加坡国家研究基金会;

关键词：

DNA nanostructures; atomic force microscopy; cryogenic transmission electron microscopy; fluorescence microscopy; spectroscopy; ELECTRON-MICROSCOPY; RADIATION-DAMAGE; 3-DIMENSIONAL RECONSTRUCTION; CONFORMATIONAL FLEXIBILITY; CRYOELECTRON MICROSCOPY; HYDRATED SPECIMENS; FOLDING DNA; RESOLUTION; CLASSIFICATION; CRYOMICROSCOPY;

D O I：

10.1002/jemt.20962

中图分类号：

R602 [外科病理学、解剖学]; R32 [人体形态学];

学科分类号：

100101 ;

摘要：

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 Forster/fluorescence resonance energy transfer (FRET) spectroscopy. Microsc. Res. Tech. 74: 688-698, 2011. (C) 2010 Wiley-Liss, Inc.

引用

页码：688 / 698

页数：11

共 115 条

[81] 3-DIMENSIONAL RECONSTRUCTION OF SINGLE PARTICLES FROM RANDOM AND NONRANDOM TILT SERIES [J].