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Nat Commun. 2018 Nov 26;9(1):4983. doi: 10.1038/s41467-018-07512-3.

High-speed AFM height spectroscopy reveals µs-dynamics of unlabeled biomolecules.

Author information

1
Weill Cornell Medicine, Department of Anesthesiology, 1300 York Avenue, New York, NY, 10065, USA.
2
Weill Cornell Medicine, Department of Physiology and Biophysics, 1300 York Avenue, New York, NY, 10065, USA.
3
Weill Cornell Medicine, Department of Anesthesiology, 1300 York Avenue, New York, NY, 10065, USA. sis2019@med.cornell.edu.
4
Weill Cornell Medicine, Department of Physiology and Biophysics, 1300 York Avenue, New York, NY, 10065, USA. sis2019@med.cornell.edu.

Abstract

Dynamics are fundamental to the functions of biomolecules and can occur on a wide range of time and length scales. Here we develop and apply high-speed AFM height spectroscopy (HS-AFM-HS), a technique whereby we monitor the sensing of a HS-AFM tip at a fixed position to directly detect the motions of unlabeled molecules underneath. This gives Angstrom spatial and microsecond temporal resolutions. In conjunction with HS-AFM imaging modes to precisely locate areas of interest, HS-AFM-HS measures simultaneously surface concentrations, diffusion coefficients and oligomer sizes of annexin-V on model membranes to decipher key kinetics allowing us to describe the entire annexin-V membrane-association and self-assembly process in great detail and quantitatively. This work displays how HS-AFM-HS can assess the dynamics of unlabeled bio-molecules over several orders of magnitude and separate the various dynamic components spatiotemporally.

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