Figure 1

Electron-beam patterning and dimension control of fluorescent nanostructures for super-resolution calibration (a) Schematic illustration of the patterning procedure: (i) a scanning focused electron beam (depicted in blue) is used to create nanoscale patterns in a PEG monolayer on an ITO-coated glass slide, (ii) the patterned slides are incubated in a buffer solution containing IgG anti-body linked fluorescent molecules or quantum dot nanoparticles, (iii) slides are washed with buffer and deionised water to remove non-specifically bound antibodies before drying, and (iv) inspected with a fluorescence microscope. (b,c) In the electron microscope, after step (i), patterned areas are visible as dark contrasted regions allowing direct evaluation of the patterning procedure and the pattern dimensions. Variations in grayscale contrast in both square and line pattern are indicative of the stochastic variations in local electron dose. Red arrows indicate small defects (b) in the patterned area or (c) on the PEG layer that may compromise pattern or background fluorescence respectively. Scale bars are 1 µm.