miEye and microEye: An open-source ecosystem for benchtop single-molecule localization microscopy and single-particle tracking

Abstract

Super-resolution fluorescence microscopy has emerged as a powerful tool for investigating biological structures at the nanoscale, well beyond the diffraction limit of classical optical imaging. Among super-resolution techniques, single-molecule localization microscopy achieves this by sequentially imaging and localizing individual fluorescent emitters, with methods such as DNA-PAINT reaching resolutions below 10 nm through transient binding of fluorescently labeled oligonucleotides. Such experiments place stringent demands on instrumentation: long acquisition times require active sample drift correction across all axes, multi-color imaging benefits from simultaneous dual-channel detection, and flexible software pipelines are essential for real-time feedback and custom workflows.