Reinforced optical cage systems enable drift-free single-molecule localization microscopy.

Abstract

Single-molecule localization microscopy achieves nanometer-scale resolution but is compromised by sample drift during image acquisition. Here we present reinforced optical cage systems, a novel approach that eliminates drift at its mechanical source rather than correcting it through complex image post-processing or fiducial markers. Reinforced optical cage systems employ perforated optomechanical components interconnected by tungsten-steel rods in a design proven by mechanical stability simulations. Our bench-top microscope, built with reinforced optical cage systems, demonstrated exceptional three-dimensional stability, with mean cumulative lateral drift of approximately 5 nanometers over 2 h in widefield fluorescence microscopy and 11-16 nanometers over 15 min in single-molecule localization microscopy, free from measurable axial drift. This development allows super-resolution microscopy to reach its full resolution without the necessity of sample drift correction, offering a straightforward, cost-effective, low-maintenance, and readily accessible solution to high-performance super-resolution microscopy. By addressing the fundamental issue of mechanical instability, reinforced optical cage systems enable improved precision instrumentation for the broader scientific and engineering community.