Open-source sub-nanometer stabilization system for super-resolution fluorescence microscopy

Area of Science:

• Optical microscopy
• Nanotechnology
• Biophysics

Background:

• Super-resolution fluorescence microscopy achieves 1-10 nm resolution for visualizing individual molecules.
• High precision requires rigorous sample drift control, especially for techniques like MINFLUX and RASTMIN.
• Active drift correction is essential for nanometer-scale resolution in microscopy.

Purpose of the Study:

• To present an active stabilization system for super-resolution microscopy.
• To achieve sub-nanometer precision stabilization for extended periods.
• To provide an adaptable and open-source solution for enhanced microscopy performance.

Main Methods:

• Development of a simple optical design for an active stabilization module.
• Integration of open-source control software with a user-friendly graphical interface.
• Demonstration with p-MINFLUX and RASTMIN measurements in diverse experimental setups.

Main Results:

• The stabilization system delivers sub-nanometer precision for hours.
• The system is adaptable as a module for various fluorescence microscopes.
• Measurements reached the theoretical Cramér-Rao Bound, resolving ~10 nm distances in DNA origami.

Conclusions:

• The active stabilization system significantly improves super-resolution microscopy precision.
• The open-source nature and adaptability facilitate widespread adoption.
• The system enables robust visualization of nanoscale structures in biological environments.