Fully automated multicolour structured illumination module for super-resolution microscopy with two excitation...

Area of Science:

• Biophysics
• Optical Microscopy
• Cell Biology

Background:

• High-resolution microscopy is crucial for studying dynamic intracellular processes.
• Conventional structured illumination microscopy (SIM) offers high resolution but is often expensive and complex.
• There is a need for cost-effective, accessible super-resolution techniques for live samples.

Purpose of the Study:

• To introduce openSIMMO, an open-source, automated SIM module for commercial microscopes.
• To enable dual-color, real-time super-resolution imaging using affordable components.
• To democratize access to SIM-based super-resolution microscopy.

Main Methods:

• Integration of single-mode fiber-coupled lasers and a digital micromirror device (DMD) with ImSwitch software.
• Development of a model for optimizing DMD diffraction patterns for various low-cost projectors.
• Implementation of an automated module compatible with existing commercial microscope systems.

Main Results:

• Achieved up to 1.55-fold improvement in lateral resolution compared to wide-field microscopy.
• Demonstrated dual-color excitation capability for simultaneous imaging of multiple targets.
• Provided a flexible open-source framework adaptable to various hardware and reconstruction algorithms.

Conclusions:

• openSIMMO offers a cost-effective and accessible solution for super-resolution microscopy.
• The open-source approach facilitates wider adoption and customization of SIM technology.
• Democratization of SIM microscopy enables broader research into dynamic cellular mechanisms.