Do-it-yourself guide: how to use the modern single-molecule toolkit

• Published in: Nature methods Vol. 5; no. 6; pp. 475 - 489
• Main Authors: Walter, Nils G, Huang, Cheng-Yen, Manzo, Anthony J, Sobhy, Mohamed A
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.06.2008; Nature Publishing Group
• Subjects: Animals; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biotechnology - methods; Cellular signal transduction; Equipment Design; Fluorescence; Fluorescence microscopy; Humans; Image Processing, Computer-Assisted; Ion channels; Life Sciences; Microscopes; Microscopy - methods; Microscopy, Atomic Force - methods; Microscopy, Fluorescence - methods; Models, Statistical; Molecular biology; Nanotechnology; Nanotechnology - methods; Optical Tweezers; Physiological aspects; Proteomics; Research methodology; review-article; Scientific apparatus & instruments; Sensitivity and Specificity; Software; Spectrometry, Fluorescence - methods; Spectroscopy; Spectrum analysis; United States
• ISSN: 1548-7091; 1548-7105; 1548-7105
• DOI: 10.1038/nmeth.1215

Single-molecule microscopy has evolved into the ultimate-sensitivity toolkit to study systems from small molecules to living cells, with the prospect of revolutionizing the modern biosciences. Here we survey the current state of the art in single-molecule tools including fluorescence spectroscopy, tethered particle microscopy, optical and magnetic tweezers, and atomic force microscopy. We also provide guidelines for choosing the right approach from the available single-molecule toolkit for applications as diverse as structural biology, enzymology, nanotechnology and systems biology.