Direct-laser writing for subnanometer focusing and single-molecule imaging

• DOI: 10.1038/s41467-022-28219-6
• Publication Date: 2022-02-03T11:07:29Z
• Authors: Simao Coelho; Jongho Baek; James Walsh; J. Justin Gooding; Katharina Gaus
• Keywords: Fiducial marker

Abstract Two-photon direct laser writing is an additive fabrication process that utilizes two-photon absorption of tightly focused femtosecond pulses to implement spatially controlled polymerization a liquid-phase photoresist. capable nanofabricating arbitrary three-dimensional structures with nanometer accuracy. Here, we explore for high-resolution optical microscopy by fabricating unique 3D fiducials single-molecule tracking and localization microscopy. By having control over the position architecture fiducials, improve axial discrimination demonstrate isotropic subnanometer focusing (<0.8 nm) tens micrometers using standard inverted microscope. We perform acquisitions cellular volumes, unsupervised data acquisition live-cell single-particle