A guide to building a low-cost centrifuge force microscope module for single-molecule force experiments

Area of Science:

• Biophysics
• Molecular Biology
• Biochemistry

Background:

• Single-molecule force spectroscopy is crucial for understanding biological processes.
• High cost and complexity of existing instruments limit accessibility.
• Need for affordable, user-friendly tools for single-molecule research.

Purpose of the Study:

• To present a protocol for building a low-cost Centrifuge Force Microscope (CFM).
• To enable high-throughput single-molecule force measurements.
• To democratize access to single-molecule biophysics techniques.

Main Methods:

• Construction of a CFM module using 3D printing and off-the-shelf components.
• Integration of the module into a commercial benchtop centrifuge.
• Development of software for CFM control and data analysis.
• Experimental measurement of force-dependent DNA duplex shearing.

Main Results:

• A functional CFM module can be built for under $1,000 in approximately one day.
• The CFM allows simultaneous probing of hundreds to thousands of single-molecule interactions.
• Successful measurement of force-dependent DNA shearing using the CFM.

Conclusions:

• The developed CFM protocol provides a cost-effective and accessible platform for single-molecule studies.
• This approach can significantly lower the barrier to entry for single-molecule research.
• Facilitates wider adoption of single-molecule techniques in biology and biophysics.