Supplementary Fig. 2.

The quality of SR image reconstructions, and the number of individual molecule localisations detected, is inversely related to electron contrast. a: WF, SR reconstruction representing 167,796 individual localisations, and overlaid images from a ‘balanced contrast’ cell expressing the GFP-C1 construct, a similar contrast level to that shown in Fig. 3. Electron micrographs with overlaid WF and SR images are also shown. b: Enlargements of the boxed areas in A showing overlay of the SR reconstruction to specific membranous structures with a much higher degree of precision compared to the WF image alone. Cc: WF, SR reconstruction representing 51,494 individual localisations, and overlaid images from a ‘high contrast’ cell expressing the GFP-C1 construct. The signal to noise ratio in the WF image worsens as electron contrast increases, as does the number of localisations for SR reconstruction. Electron micrographs with overlaid WF and SR images are also shown. D: Enlargements of the boxed areas in c. As before, localisation of signal to specific membranous structures is possible, though it is clear that the decreased number of localisations reduces the certainty with which signals can be assigned to specific structures. G: Golgi, M: mitochondria, V: vesicles. Scale bars – 5 µm (a, c), 500 nm (b, d).