Fully-Automated Multicolour Structured Illumination Module for Super-resolution Microscopy

<p>&nbsp;</p> <p>In the rapidly advancing field of biological imaging, high-resolution techniques that are cost-effective and accessible are essential for observing and understanding intracellular dynamics. Structured illumination microscopy (SIM) is a preferred method for achieving high axial and lateral resolution in living samples due to its optical sectioning and minimal phototoxicity. However, the high cost and complexity of conventional SIM systems limit their widespread use. In our work, we present an open-source, fully-automated, two-color structured illumination module that is compatible with commercially available microscope stands. The compact design, which includes low-cost single-mode fiber-coupled lasers and a digital micromirror device (DMD), is integrated into the open-source acquisition and control software ImSwitch to facilitate real-time super-resolution imaging. This system achieves up to a 1.55-fold improvement in lateral resolution compared to conventional wide-field microscopy.&nbsp;</p> <p>To ensure optimal DMD diffraction performance, we developed a model using tilt and roll pixels, enabling the use of low-cost video projectors in coherent SIM setups. Our aim is to democratize SIM-based super-resolution microscopy by providing comprehensive open-source documentation and a modular software framework compatible with various hardware components (e.g., cameras, stages) and reconstruction algorithms.&nbsp;</p> <p>All datasets generated and analyzed during this study are openly available and can be accessed through our public repository <a href="https://opensimmo.github.io/">[repository link]</a>. The datasets include raw and processed images, calibration files, and software scripts, enabling replication and further innovation. This approach will help upgrade as many devices as possible to the super-resolution realm, fostering greater accessibility and collaboration in the scientific community</p>