High-Yield Characterization of Single Molecule Interactions with DeepTip^TM Atomic Force Microscopy Probes

Area of Science:

• Biophysics
• Surface Science
• Nanotechnology

Background:

• Characterizing single molecule interactions is crucial for understanding biological processes.
• Existing methodologies often face challenges with yield and specificity.
• Atomic force microscopy (AFM) offers high-resolution force measurements.

Purpose of the Study:

• To develop a comprehensive methodology for high-yield identification and analysis of single molecular events.
• To establish a robust system for studying molecular interactions using functionalized probes.
• To demonstrate the versatility of the proposed technique for various molecular systems.

Main Methods:

• Utilized functionalized DeepTip™ probes for single molecule force spectroscopy.
• Employed biotin-streptavidin interactions as a model system.
• Developed a procedure involving covalent binding of target and sensing molecules to surfaces and probes, respectively.
• Acquired force-displacement curves using atomic force microscopy.

Main Results:

• Achieved molecular resolution consistently in single molecule force measurements.
• Demonstrated a success rate higher than 90% for identified molecular events.
• Observed a very low level of unspecific interactions, indicating high specificity.
• Validated the robustness and versatility of the developed methodology.

Conclusions:

• The proposed methodology provides a high-yield and specific approach for analyzing single molecule interactions.
• The technique is robust and versatile, suitable for a wide range of molecular studies.
• DeepTip™ probes combined with AFM offer a powerful platform for nanoscale biophysical investigations.