title: Tuning the Music: Acoustic Force Spectroscopy (AFS) 2.0.
abstract: AFS is a recently introduced high-throughput single-molecule technique that allows studying structural and mechanochemical properties of many biomolecules in parallel. To further improve the method, we developed a modelling tool to optimize the layer thicknesses, and a calibration method to experimentally validate the modelled force profiles. After optimization, we are able to apply 350pN on 4.5μm polystyrene beads, without the use of an amplifier, at the coverslip side of the AFS chip. Furthermore, we present the use of a transparent piezo to generate the acoustic force and we show that AFS can be combined with high-NA oil or water-immersion objectives. With this set of developments AFS will be applicable to a broad range of single-molecule experiments.
authors.affiliation: Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands; LaserLaB Amsterdam, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
authors.forename: Douwe
authors.initials: D
authors.lastname: Kamsma
authors.affiliation: Department of Physics and Astronomy, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Ramon
authors.initials: R
authors.lastname: Creyghton
authors.affiliation: Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands; LaserLaB Amsterdam, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands; LUMICKS B.V., Amsterdam, The Netherlands.
authors.forename: Gerrit
authors.initials: G
authors.lastname: Sitters
authors.affiliation: Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands; LaserLaB Amsterdam, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands. Electronic address: g.j.l.wuite@vu.nl.
authors.forename: Gijs J L
authors.initials: GJ
authors.lastname: Wuite
authors.affiliation: Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands; LaserLaB Amsterdam, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
authors.forename: Erwin J G
authors.initials: EJ
authors.lastname: Peterman
mesh_terms: D000162:Acoustics / Q000295:instrumentation*; D055595:Mechanical Phenomena; D018625:Microscopy, Atomic Force / Q000379:methods*; D009696:Nucleic Acids / Q000737:chemistry / Q000302:isolation & purification*; D000072760:Single Molecule Imaging / Q000379:methods*
chemical_list: D009696:Nucleic Acids
keywords: AFS; Acoustic force spectroscopy; DNA tethering; Layered resonator; Transparent piezo