Fig. 1: DNA-PAINT imaging of DNA origami structures on an SDC-OPR microscope.
a Schematic of the Spinning Disk Confocal with Optical Photon Reassignment (SDC-OPR) system (CSU-W1 SoRA Nikon), featuring a microlens array designed to minimize pinhole size and maximize photon collection. b DNA origami design with six DNA-PAINT docking strands arranged in three pairs: 53 nm apart overall; central pair exhibiting the largest separation of 17 nm, while the edge pairs are spaced 10 nm apart. Detailed sample preparation and imaging conditions are provided in the Methods section and Supplementary Table 1. Staple modifications can be found in Supplementary Table 2. c Spinning Disk Confocal (SDC) DNA-PAINT image of a representative DNA origami (top), and position distribution for the left (top) and central (bottom) docking strands pairs, showing measured distances. d SDC-OPR DNA-PAINT image of a representative DNA origami (top), and position distribution of the left (top) and central (bottom) docking pairs, showing measured distances. e DNA origami design featuring 6 nm-spaced docking sites to assess achievable resolution (top). Staple modifications can be found in Supplementary Table 3. DNA-PAINT image of a representative DNA origami (central), and position distribution for the docking pair, highlighting measured distance (bottom). f Localization precision (σSMLM) across three different FOVs sizes enabled by the SDC-OPR system: 53 µm, 76 µm and 211 µm side size. Representative super-resolved DNA origami images corresponding to each FOV size are shown in the three top panels. The bottom panel displays the distribution of localization precision (σSMLM) as histograms, with different colors for each magnification. All DNA origami sample imaging were repeated in two independent experiments. Source data of histograms is provided as a Source Data file. Panel (a) and DNA origami sketches of panel (b) and (e) were created with BioRender.com.
