title: Multiplexed single-molecule force spectroscopy using a centrifuge.
abstract: We present a miniature centrifuge force microscope (CFM) that repurposes a benchtop centrifuge for high-throughput single-molecule experiments with high-resolution particle tracking, a large force range, temperature control and simple push-button operation. Incorporating DNA nanoswitches to enable repeated interrogation by force of single molecular pairs, we demonstrate increased throughput, reliability and the ability to characterize population heterogeneity. We perform spatiotemporally multiplexed experiments to collect 1,863 bond rupture statistics from 538 traceable molecular pairs in a single experiment, and show that 2 populations of DNA zippers can be distinguished using per-molecule statistics to reduce noise.
authors.affiliation: School of engineering and applied sciences, Harvard University, Cambridge, Massachusetts 02138, USA.
authors.forename: Darren
authors.identifier: http://orcid.org/0000-0002-2271-6910
authors.initials: D
authors.lastname: Yang
authors.affiliation: Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Ward
authors.affiliation: Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.|The RNA Institute, University at Albany, Albany, New York 12222, USA.
authors.forename: Ken
authors.identifier: http://orcid.org/0000-0002-2578-1339
authors.initials: K
authors.lastname: Halvorsen
authors.affiliation: Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, Massachusetts 02115, USA.|Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.|Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA.
authors.forename: Wesley P
authors.initials: WP
authors.lastname: Wong
mesh_terms: D002498:Centrifugation / Q000295:instrumentation*; D004247:DNA / Q000737:chemistry; D018625:Microscopy, Atomic Force / Q000379:methods*; D009690:Nucleic Acid Conformation; D013057:Spectrum Analysis; D013696:Temperature
chemical_list: D004247:DNA