  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.
