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bioRxiv · 10.64898/2026.03.10.710894

Open Blink: Low-cost TIRF microscopy for super-resolutionimaging via μManager

Huo, R.·Komen, J.·Engelhardt, M. L. K.·Millot, A.·Extermann, J.·Grussmayer, K.
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Abstract

Super-resolution localization microscopy (SMLM) has become a central tool for nanoscale biological research for its high spatial resolution and compatibility with wide-field microscopy. Achieving quantitative SMLM, however, requires homogeneous high-power illumination, nanometric axial stability, and precise multi-channel detection, features typically restricted to high-end commercial instruments or custom solutions in specialized laboratories. The cost of such microscopes and their technical complexity still limit the accessibility of these advanced imaging techniques. Several home-made single molecule microscopes and their submodules have been demonstrated as opensource, highly-customizable, and cost-effective alternatives for their commercial counterparts. Yet, implementation of such systems often requires expert knowledge in optics, electronics, and control system engineering. We introduce Open Blink, a compact open-source TIRF microscope integrating powerful homogeneous quad-line laser illumination, dual-channel detection, and active focus-lock stabilization for quantitative multi-color super-resolution imaging. Open Blink achieves a localization precision below 10 nm in dSTORM, supports a tunable, large field of view from 105 x 105 {micro}m2 up to 257 x 257 {micro}m2, and maintains axial stability over hours, enabling high-throughput super-resolution acquisition. Built with predominantly off-the-shelf components, and full integration into the open-source software {micro}Manager where metadata registration ensures reproducibility, Open Blink offers a low threshold for adoption by easing implementation, use and maintenance. At a substantially reduced cost of approximately 70 000 Euros, among which the high-power laser combiner alone is less than 20 000 euros, Open Blink greatly improves accessibility for laboratories who wish to implement scalable high performance super-resolution microscopy based on single molecules.

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Huo, R., Komen, J., Engelhardt, M. L. K., Millot, A., Extermann, J., Grussmayer, K.. 2026-03-13. Open Blink: Low-cost TIRF microscopy for super-resolutionimaging via μManager. https://doi.org/10.64898/2026.03.10.710894

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