Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 2001 Nov;81(5):2946–2953. doi: 10.1016/S0006-3495(01)75934-6

Force measurements on single molecular contacts through evanescent wave microscopy.

G Zocchi 1
PMCID: PMC1301758  PMID: 11606304

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.

Full Text

The Full Text of this article is available as a PDF (131.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allen S., Davies J., Davies M. C., Dawkes A. C., Roberts C. J., Tendler S. J., Williams P. M. The influence of epitope availability on atomic-force microscope studies of antigen-antibody interactions. Biochem J. 1999 Jul 1;341(Pt 1):173–178. [PMC free article] [PubMed] [Google Scholar]
  2. Allen S., Davies J., Dawkes A. C., Davies M. C., Edwards J. C., Parker M. C., Roberts C. J., Sefton J., Tendler S. J., Williams P. M. In situ observation of streptavidin-biotin binding on an immunoassay well surface using an atomic force microscope. FEBS Lett. 1996 Jul 22;390(2):161–164. doi: 10.1016/0014-5793(96)00651-5. [DOI] [PubMed] [Google Scholar]
  3. Block S. M., Goldstein L. S., Schnapp B. J. Bead movement by single kinesin molecules studied with optical tweezers. Nature. 1990 Nov 22;348(6299):348–352. doi: 10.1038/348348a0. [DOI] [PubMed] [Google Scholar]
  4. Boland T., Ratner B. D. Direct measurement of hydrogen bonding in DNA nucleotide bases by atomic force microscopy. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5297–5301. doi: 10.1073/pnas.92.12.5297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cluzel P., Lebrun A., Heller C., Lavery R., Viovy J. L., Chatenay D., Caron F. DNA: an extensible molecule. Science. 1996 Feb 9;271(5250):792–794. doi: 10.1126/science.271.5250.792. [DOI] [PubMed] [Google Scholar]
  6. Coppin C. M., Pierce D. W., Hsu L., Vale R. D. The load dependence of kinesin's mechanical cycle. Proc Natl Acad Sci U S A. 1997 Aug 5;94(16):8539–8544. doi: 10.1073/pnas.94.16.8539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Drake B., Prater C. B., Weisenhorn A. L., Gould S. A., Albrecht T. R., Quate C. F., Cannell D. S., Hansma H. G., Hansma P. K. Imaging crystals, polymers, and processes in water with the atomic force microscope. Science. 1989 Mar 24;243(4898):1586–1589. doi: 10.1126/science.2928794. [DOI] [PubMed] [Google Scholar]
  8. Essevaz-Roulet B., Bockelmann U., Heslot F. Mechanical separation of the complementary strands of DNA. Proc Natl Acad Sci U S A. 1997 Oct 28;94(22):11935–11940. doi: 10.1073/pnas.94.22.11935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Evans E., Ritchie K. Dynamic strength of molecular adhesion bonds. Biophys J. 1997 Apr;72(4):1541–1555. doi: 10.1016/S0006-3495(97)78802-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Florin E. L., Moy V. T., Gaub H. E. Adhesion forces between individual ligand-receptor pairs. Science. 1994 Apr 15;264(5157):415–417. doi: 10.1126/science.8153628. [DOI] [PubMed] [Google Scholar]
  11. Hinterdorfer P., Baumgartner W., Gruber H. J., Schilcher K., Schindler H. Detection and localization of individual antibody-antigen recognition events by atomic force microscopy. Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3477–3481. doi: 10.1073/pnas.93.8.3477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Howard J., Hudspeth A. J., Vale R. D. Movement of microtubules by single kinesin molecules. Nature. 1989 Nov 9;342(6246):154–158. doi: 10.1038/342154a0. [DOI] [PubMed] [Google Scholar]
  13. Kellermayer M. S., Smith S. B., Granzier H. L., Bustamante C. Folding-unfolding transitions in single titin molecules characterized with laser tweezers. Science. 1997 May 16;276(5315):1112–1116. doi: 10.1126/science.276.5315.1112. [DOI] [PubMed] [Google Scholar]
  14. Kojima H., Muto E., Higuchi H., Yanagida T. Mechanics of single kinesin molecules measured by optical trapping nanometry. Biophys J. 1997 Oct;73(4):2012–2022. doi: 10.1016/S0006-3495(97)78231-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Löster K., Horstkorte R. Enzymatic quantification of cell-matrix and cell-cell adhesion. Micron. 2000 Jan;31(1):41–53. doi: 10.1016/s0968-4328(99)00062-1. [DOI] [PubMed] [Google Scholar]
  16. Merkel R., Nassoy P., Leung A., Ritchie K., Evans E. Energy landscapes of receptor-ligand bonds explored with dynamic force spectroscopy. Nature. 1999 Jan 7;397(6714):50–53. doi: 10.1038/16219. [DOI] [PubMed] [Google Scholar]
  17. Meyhöfer E., Howard J. The force generated by a single kinesin molecule against an elastic load. Proc Natl Acad Sci U S A. 1995 Jan 17;92(2):574–578. doi: 10.1073/pnas.92.2.574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Noji H., Yasuda R., Yoshida M., Kinosita K., Jr Direct observation of the rotation of F1-ATPase. Nature. 1997 Mar 20;386(6622):299–302. doi: 10.1038/386299a0. [DOI] [PubMed] [Google Scholar]
  19. Radmacher M., Fritz M., Hansma H. G., Hansma P. K. Direct observation of enzyme activity with the atomic force microscope. Science. 1994 Sep 9;265(5178):1577–1579. doi: 10.1126/science.8079171. [DOI] [PubMed] [Google Scholar]
  20. Rief M., Gautel M., Oesterhelt F., Fernandez J. M., Gaub H. E. Reversible unfolding of individual titin immunoglobulin domains by AFM. Science. 1997 May 16;276(5315):1109–1112. doi: 10.1126/science.276.5315.1109. [DOI] [PubMed] [Google Scholar]
  21. Singh-Zocchi M., Andreasen A., Zocchi G. Osmotic pressure contribution of albumin to colloidal interactions. Proc Natl Acad Sci U S A. 1999 Jun 8;96(12):6711–6715. doi: 10.1073/pnas.96.12.6711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Smith S. B., Cui Y., Bustamante C. Overstretching B-DNA: the elastic response of individual double-stranded and single-stranded DNA molecules. Science. 1996 Feb 9;271(5250):795–799. doi: 10.1126/science.271.5250.795. [DOI] [PubMed] [Google Scholar]
  23. Smith S. B., Finzi L., Bustamante C. Direct mechanical measurements of the elasticity of single DNA molecules by using magnetic beads. Science. 1992 Nov 13;258(5085):1122–1126. doi: 10.1126/science.1439819. [DOI] [PubMed] [Google Scholar]
  24. Svoboda K., Block S. M. Force and velocity measured for single kinesin molecules. Cell. 1994 Jun 3;77(5):773–784. doi: 10.1016/0092-8674(94)90060-4. [DOI] [PubMed] [Google Scholar]
  25. Svoboda K., Schmidt C. F., Schnapp B. J., Block S. M. Direct observation of kinesin stepping by optical trapping interferometry. Nature. 1993 Oct 21;365(6448):721–727. doi: 10.1038/365721a0. [DOI] [PubMed] [Google Scholar]
  26. Tskhovrebova L., Trinick J., Sleep J. A., Simmons R. M. Elasticity and unfolding of single molecules of the giant muscle protein titin. Nature. 1997 May 15;387(6630):308–312. doi: 10.1038/387308a0. [DOI] [PubMed] [Google Scholar]
  27. Vale R. D., Funatsu T., Pierce D. W., Romberg L., Harada Y., Yanagida T. Direct observation of single kinesin molecules moving along microtubules. Nature. 1996 Apr 4;380(6573):451–453. doi: 10.1038/380451a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wang M. D., Schnitzer M. J., Yin H., Landick R., Gelles J., Block S. M. Force and velocity measured for single molecules of RNA polymerase. Science. 1998 Oct 30;282(5390):902–907. doi: 10.1126/science.282.5390.902. [DOI] [PubMed] [Google Scholar]
  29. Wong J., Chilkoti A., Moy V. T. Direct force measurements of the streptavidin-biotin interaction. Biomol Eng. 1999 Dec 31;16(1-4):45–55. doi: 10.1016/s1050-3862(99)00035-2. [DOI] [PubMed] [Google Scholar]
  30. Yasuda R., Noji H., Kinosita K., Jr, Yoshida M. F1-ATPase is a highly efficient molecular motor that rotates with discrete 120 degree steps. Cell. 1998 Jun 26;93(7):1117–1124. doi: 10.1016/s0092-8674(00)81456-7. [DOI] [PubMed] [Google Scholar]
  31. Yin H., Wang M. D., Svoboda K., Landick R., Block S. M., Gelles J. Transcription against an applied force. Science. 1995 Dec 8;270(5242):1653–1657. doi: 10.1126/science.270.5242.1653. [DOI] [PubMed] [Google Scholar]
  32. Zocchi G. Proteins unfold in steps. Proc Natl Acad Sci U S A. 1997 Sep 30;94(20):10647–10651. doi: 10.1073/pnas.94.20.10647. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES