Force measurements on single molecular contacts through evanescent wave microscopy

Biophys J. 2001 Nov;81(5):2946-53. doi: 10.1016/S0006-3495(01)75934-6.

Abstract

We introduce a new method to apply controlled forces on single molecules. The motion of a micron-sized bead attached to a solid surface through a single molecular contact is tracked by evanescent wave microscopy as a force is exerted through a flow. We report measurements of the streptavidin-biotin bond rupture force obtained with this technique. We also obtain detailed measurements of the balance of forces involved in detaching an adhering bead with a flow. A small lateral force translates into a much bigger normal force on the attachment point. This effect is relevant for the interpretation of common cell adhesion assays.

MeSH terms

  • Biological Factors / chemistry
  • Biotin / chemistry*
  • Cell Adhesion Molecules / chemistry
  • Microscopy / methods*
  • Microscopy, Atomic Force / methods
  • Streptavidin / chemistry*

Substances

  • Biological Factors
  • Cell Adhesion Molecules
  • Biotin
  • Streptavidin