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Grabbing the cat by the tail: manipulating molecules one by one

Nature Reviews Molecular Cell Biology volume 1, pages 130–136 (2000)Cite this article

Abstract

Methods for manipulating single molecules are yielding new information about both the forces that hold biomolecules together and the mechanics of molecular motors. We describe here the physical principles behind these methods, and discuss their capabilities and current limitations.

Key Points

  • Methods for manipulating single molecules are yielding new information about both the forces that hold biomolecules together and the mechanics of molecular motors.

  • All single-molecule manipulation methods require two basic elements: a probe, which is usually of microscopic dimensions, that can generate or detect forces and displacements; and a way to spatially locate the molecules.

  • Mechanical force transducers apply or sense forces through the displacement of a bendable beam. The most common examples are SFM cantilevers and microneedles.

  • The advantages of SFM are its high spatial range and sensitivity, its throughput (the ability to study many single molecules on a surface) and versatility.

  • Glass microneedles are usually softer than SFM cantilevers, giving them an advantage for probing delicate biological systems.

  • Unlike mechanical transducers, external fields act on molecules from a distance. These fields can be used to exert forces on molecules by acting either on the molecules themselves, or through 'handles' such as glass beads, polystyrene beads or metallic particles attached to the molecules.

  • Flow fields exert forces on objects through the transfer of momentum from the fluid to the object.

  • Magnetic fields can be used to manipulate and apply very stable and small forces to biomolecules that are tethered to magnetic particles.

  • Optical tweezers use radiation pressure (which stems from the momentum change as light refracts off an object) to hold objects in a focused laser beam, with which it is possible to generate a spring-like force.

  • The future of biomolecular manipulation depends on three factors: the integration and further development of single molecule techniques, progress in the field of nanotechnology, and the use of high-throughput systems such as microfluidics (microscopic liquid channels).

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Figure 1: Applications of the scanning force microscope (SFM).
Figure 2: Using a microneedle to measure the force of myosin acting on actin.
Figure 3: Geometries of typical single-molecule experiments.
Figure 4: Geometries of typical optical-trap single-molecule experiments.

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Acknowledgements

We thank S. Smith and J. Choy for their helpful comments. This work was supported in part by grants from the NIH and the NSF (to C.B.).

Author information

Authors and Affiliations

  1. Howard Hughes Medical Institute, University of California , Berkeley, 94720, California, USA

    Carlos Bustamante

  2. Departments of Molecular and Cell Biology, University of California, Berkeley, 94720, California, USA

    Carlos Bustamante & Jed C. Macosko

  3. Physics, University of California, Berkeley , 94720, California, USA

    Carlos Bustamante & Gijs J. L. Wuite

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  1. Carlos Bustamante
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  2. Jed C. Macosko
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  3. Gijs J. L. Wuite
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Related links

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FURTHER INFORMATION

SFM overview

SFM in depth

Microneedle research page

Movies of flow fields stretching DNA

Theory of optical tweezers

Building optical tweezers

Background of optical tweezers

Microfluidics applications

Microfluidics applications

Glossary

OPTICAL TWEEZERS

Focused photon fields.

PIEZO-ELECTRIC

Describes a device that expands or contracts as a voltage is applied to an internal crystal.

HYDRODYNAMIC FIELD

A force field resulting from the momentum imparted by molecules in a flowing aqueous solution.

PHOTON FIELD

A force field resulting from the momentum imparted by photons in a beam of light.

LAMINAR FLOW

A flow of molecules in which neighbouring molecules have linearly dependent velocities, that is, not a turbulent flow.

STOKES'S LAW

 Fdrag = 6πrη v

RADIATION PRESSURE

The pressure on an object that arises from photon collisions rather than from bombarding molecules.

NANOTECHNOLOGY

Any technological development that exceeds standard lower size limits of modern microfabrication techniques (hundreds of nanometres or less).

MICROFLUIDICS

Microscopic channels etched into a surface by modern microfabrication techniques for the purpose of transporting small amounts of solution from one place to another.

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Bustamante, C., Macosko, J. & Wuite, G. Grabbing the cat by the tail: manipulating molecules one by one. Nat Rev Mol Cell Biol 1, 130–136 (2000). https://doi.org/10.1038/35040072

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