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Updated: Mar 21, 2026

Force Spectroscopy of Single Protein Molecules Using an Atomic Force Microscope
Published on: February 28, 2019
Tuning the Music: Acoustic Force Spectroscopy (AFS) 2.0
Douwe Kamsma1, Ramon Creyghton2, Gerrit Sitters3
1Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands; LaserLaB Amsterdam, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
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.
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