Massively parallel single-molecule manipulation using centrifugal force

TL;DR

This paper introduces the Centrifuge Force Microscope (CFM), a novel instrument enabling massively parallel, high-throughput single-molecule force measurements using centrifugal force, significantly reducing time and cost compared to traditional methods.

Contribution

The authors developed a new centrifuge-based instrument for parallel single-molecule force spectroscopy, allowing rapid, cost-effective, and versatile measurements without calibration.

Findings

Performed thousands of rupture experiments in parallel

Measured DNA overstretching transition at 66 ± 3 pN

Demonstrated rapid characterization of unbinding kinetics

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 with a new approach: massively parallel single-molecule force measurements using centrifugal force. This approach is realized in a novel instrument that we call the Centrifuge Force Microscope (CFM), 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 $\pm$ 3 pN. With significant benefits in efficiency, cost, simplicity, and versatility, "single-molecule centrifugation" has the potential to revolutionize single-molecule experimentation, and open access to a wider range of researchers and experimental systems.