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High-Speed Atomic Force Microscopy

High-Speed Atomic Force Microscopy (HS-AFM Ando model) is a powerful visualization technique that allows simultaneous acquisition of the structural and kinetic parameters of the single molecules or molecular assemblies at the nanoscale spatial resolution and millisecond temporal resolution.

The important features of the HS-AFM:AFM-image

Mirror tilting system; Compact and lightweight AFM head; High-speed feedback control;

Ultra-small cantilever; Integrated with optical microscope; Selection of an observation area;

Various size of samples.

The advantages of HS-AFM used by us in various projects related to the different aspects of muscle contraction:

Thin filaments activation (see here for details: Matusovsky et al., 2019);

Nano-mechanics and force-generating mechanism of skeletal myosin II in actin-myosin complex (see here for details: Matusovsky et al., 2021);

Cooperativity between the myosin motors (the paper is under review).

(a) A schematic view of a HS-AFM set-up. A surface of the sample stage is scanned in solution by a small and sharp tip attached to a soft cantilever. A laser beam is focused on the backside of the cantilever and captures the topography of the sample.

(b) HS-AFM image of the double-headed proteolytic fragment of skeletal myosin II (HMM), where two of each myosin head is bound between two parallel actin filaments. Two globular heads and the lever arms of HMM are visible.

(c) Corresponding structural model of myosin single-headed S1 fragment that showed certain lever arm configurations. The lower and upper S1 structures represent a pre-power stroke (PDB: 1QVI) and myosin-ADP (PDB: 3I5F)states, respectively.

 

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