Quantitative guidelines for force calibration through spectral analysis of magnetic tweezers data

Aartjan J.W. Te Velthuis, Jacob W.J. Kerssemakers, Jan Lipfert, Nynke H. Dekker

Research output: Contribution to journalArticlepeer-review

68 Scopus citations

Abstract

Single-molecule techniques are powerful tools that can be used to study the kinetics and mechanics of a variety of enzymes and their complexes. Force spectroscopy, for example, can be used to control the force applied to a single molecule and thereby facilitate the investigation of real-time nucleic acid-protein interactions. In magnetic tweezers, which offer straightforward control and compatibility with fluorescence measurements or parallel tracking modes, force-measurement typically relies on the analysis of positional fluctuations through video microscopy. Significant errors in force estimates, however, may arise from incorrect spectral analysis of the Brownian motion in the magnetic tweezers. Here we investigated physical and analytical optimization procedures that can be used to improve the range over which forces can be reliably measured. To systematically probe the limitations of magnetic tweezers spectral analysis, we have developed a magnetic tweezers simulator, whose outcome was validated with experimental data. Using this simulator, we evaluate methods to correctly perform force experiments and provide guidelines for correct force calibration under configurations that can be encountered in typical magnetic tweezers experiments.

Original languageEnglish (US)
Pages (from-to)1292-1302
Number of pages11
JournalBiophysical Journal
Volume99
Issue number4
DOIs
StatePublished - Aug 9 2010
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biophysics

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