Minimizing pulling geometry errors in atomic force microscope single molecule force spectroscopy

Biophys J. 2008 Oct;95(8):3991-8. doi: 10.1529/biophysj.108.138842. Epub 2008 Jul 18.

Abstract

In atomic force microscopy-based single molecule force spectroscopy (AFM-SMFS), it is assumed that the pulling angle is negligible and that the force applied to the molecule is equivalent to the force measured by the instrument. Recent studies, however, have indicated that the pulling geometry errors can drastically alter the measured force-extension relationship of molecules. Here we describe a software-based alignment method that repositions the cantilever such that it is located directly above the molecule's substrate attachment site. By aligning the applied force with the measurement axis, the molecule is no longer undergoing combined loading, and the full force can be measured by the cantilever. Simulations and experimental results verify the ability of the alignment program to minimize pulling geometry errors in AFM-SMFS studies.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Artifacts*
  • Computer Simulation
  • DNA, Viral / chemistry
  • Dextrans / chemistry
  • Microscopy, Atomic Force*
  • Software
  • Spectrum Analysis / methods*
  • Surface Properties

Substances

  • DNA, Viral
  • Dextrans