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Review

Nature Methods 5, 475–489 (1 June 2008) | doi:10.1038/nmeth.1215

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

Nils G Walter , Cheng-Yen Huang , Anthony J Manzo & Mohamed A Sobhy

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.