Single-Molecule Manipulation for the Masses: New Technique Offers Dramatic Improvements in Throughput and Cost
ScienceDaily (June 2, 2010) — Scientists have developed a new massively-parallel approach for manipulating single DNA and protein molecules and studying their interactions under force.
This is the Centrifuge Force Micoscope in action. (Credit: Dr. Wesley Wong)
The finding appears in the June 2 issue of Biophysical Journal.
The team of researchers from the Rowland Institute at Harvard University claim that their technique, which they call "single molecule centrifugation," offers dramatic improvements in throughput and cost compared with more established techniques.
"By combining a microscope and a centrifuge, forces can be applied to many molecules at once while simultaneously observing their nano-to-microscale motions," explains author Wesley P. Wong, a Principal Investigator at Rowland.
Recent technologies such as optical and magnetic tweezers and the Atomic Force Microscope (AFM) have enabled the mechanical manipulation of single molecules, leading to new insights in biological systems ranging from DNA replication to blood clotting.
...
+++++
Copyright © 2010 Biophysical Society. All rights reserved.
Biophysical Journal, Volume 98, Issue 11, L53-L55, 2 June 2010
doi:10.1016/j.bpj.2010.03.012
Massively Parallel Single-Molecule Manipulation Using Centrifugal Force
Ken Halvorsen and Wesley P. Wong,
The Rowland Institute at Harvard, Harvard University, Cambridge, Massachusetts
Abstract
Precise manipulation of single molecules has already led to remarkable insights in physics, chemistry, biology, and medicine. However, widespread adoption of single-molecule techniques has been impeded by equipment cost and the laborious nature of making measurements one molecule at a time. We have solved these issues by developing an approach that enables massively parallel single-molecule force measurements using centrifugal force. This approach is realized in an instrument that we call the centrifuge force microscope in which objects in an orbiting sample are subjected to a calibration-free, macroscopically uniform force-field while their micro-to-nanoscopic motions are observed. We demonstrate high-throughput single-molecule force spectroscopy with this technique by performing thousands of rupture experiments in parallel, characterizing force-dependent unbinding kinetics of an antibody-antigen pair in minutes rather than days. Additionally, we verify the force accuracy of the instrument by measuring the well-established DNA overstretching transition at 66 ± 3 pN. With significant benefits in efficiency, cost, simplicity, and versatility, single-molecule centrifugation has the potential to expand single-molecule experimentation to a wider range of researchers and experimental systems.
+++++
+++++
