An ultra-wide scanner for large-area high-speed atomic force microscopy with megapixel resolution

Area of Science:

• Biophysics
• Nanotechnology
• Materials Science

Background:

• High-speed atomic force microscopy (HS-AFM) excels at visualizing individual biomolecule dynamics.
• Current HS-AFM systems have limited scan ranges (hundreds of nanometers), hindering the study of larger molecular assemblies like protein crystals or aggregates.
• Existing large-scan HS-AFM designs often compromise speed, resolution, or stability.

Purpose of the Study:

• To develop a novel HS-AFM sample-scanner system capable of imaging large areas with high resolution and speed.
• To overcome the limitations of conventional HS-AFM for observing large-scale dynamic molecular processes.
• To enable detailed analysis of complex biological samples and molecular arrays.

Main Methods:

• Development of a flexure-based HS-AFM sample-scanner system.
• Implementation of a high-speed scanning capability (up to 7.2 mm/s) with a high resonance frequency (>2 kHz).
• Acquisition of large topographic images (up to 36x36 µm²) with up to 16 megapixels and molecular resolution.

Main Results:

• The new system successfully records large-area (≤36x36 µm²) megapixel topographic images with consistent nanometer resolution.
• Stable operation at high scan speeds (7.2 mm/s) was achieved, significantly reducing imaging time.
• Time-lapse mode enabled simultaneous observation of 2D crystal growth, domain orientation, and lattice defects in Annexin A5 crystals.

Conclusions:

• The developed HS-AFM sample-scanner system expands imaging capabilities to large dynamic molecular arrays and complex biological surfaces.
• It bridges the gap between single-molecule imaging and the study of larger biological structures at high resolution.
• This technology facilitates quantitative analysis of heterogeneous samples, including cellular surfaces, within biologically relevant time frames.