Force-Fluorescence Spectroscopy at the Single-Molecule Level

Ruobo Zhou, Michael Schlierf, Taekjip Ha

Research output: Chapter in Book/Report/Conference proceedingChapter

23 Scopus citations

Abstract

During the past decade, various powerful single-molecule techniques have evolved and helped to address important questions in life sciences. Yet these techniques would be even more powerful if they would be combined, that is, single-molecule manipulation with an orthogonal single-molecule observation. Here, we present a recently developed approach to combine single-molecule optical tweezers with single-molecule fluorescence spectroscopy. Optical tweezers are used to manipulate and observe mechanical properties on the nanometer scale and piconewton force range. However, once the force range is in the low piconewton range or less, the spatial resolution of optical tweezers decreases significantly. In combination with fluorescence spectroscopy, like Förster resonance energy transfer (FRET), we are able to observe nanometer fluctuations and internal conformational changes in a low-force regime. The possibility to place fluorescent labels at nearly any desired position and a sophisticated design of the experiment increases the amount of information that can be extracted in contrast to pure mechanical or fluorescence experiments.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages405-426
Number of pages22
EditionC
DOIs
StatePublished - 2010

Publication series

NameMethods in Enzymology
NumberC
Volume475
ISSN (Print)0076-6879

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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