DNA-PAINT Probe Modifications Support High-Resolution Imaging with Shorter Binding Domains

Area of Science:

• Nanotechnology
• Microscopy
• Molecular Biology

Background:

• DNA-based Points Accumulation for Imaging in Nanoscale Topography (DNA-PAINT) is a powerful super-resolution technique.
• Optimizing DNA-PAINT is essential for advancing nanoscale detection and metrology.
• Current methods often rely on longer probes, limiting detection density.

Purpose of the Study:

• To develop modified imager probes for enhanced DNA-PAINT performance.
• To enable imaging of shorter DNA sequences with improved stability and resolution.
• To advance applications in nanoscience and nanotechnology, such as Nucleic Acid Memory.

Main Methods:

• Introduced structural and chemical modifications to imager probes, including mini-hairpins and Bridged Nucleic Acids (BNA).
• These modifications increase the thermomechanical stability of DNA duplexes.
• Validated probe performance using DNA nanostructure devices for controlled in vitro testing.

Main Results:

• Achieved DNA-PAINT imaging with approximately 5 nm resolution using a 4-nucleotide hybridization domain, 43% shorter than previous probes.
• Demonstrated equivalent imaging resolution compared to state-of-the-art DNA-PAINT probes.
• Probes facilitate targeting more binding domains per localization unit, increasing information density.

Conclusions:

• Modified imager probes significantly enhance DNA-PAINT capabilities for imaging shorter DNA domains.
• This strategy is vital for high-density information storage in applications like Nucleic Acid Memory.
• The developed probes offer a pathway for improved nanoscale metrology and nanoscience research.