Extended Data Fig. 5: Super-resolution imaging of a living cell presenting tubulin bundles within a cultured neuron with SIM and eSRRF.

a The microtubule network in a cultured neuron expressing Tubulin-eGFP can be visualized with SIM at a resolution level (WF-FRC: 253 ± 74 nm, SIM-FRC: 202 ± 47 nm) which allows to resolve microtubule bundles in single dendrites. b In cultured neuron expressing Skylan-NS-Tubulin imaged using TIRFM successful eSRRF processing requires user intervention in eSRRF parameter selection due to the local variability and nonlinearity of the QnR map. Especially in large field-of-view images, distinct structural features and variability in marker density result in different optimal parameters for eSRRF super-resolution processing. For example, in this 100 µm × 60 µm image of the microtubule network regions i & ii display a QnR maximum (marked by the red square) at different positions within the parameter sweep map (left to right: R = 3.0–7.5 in steps of 0.5, top to bottom: S = 1–6 in steps of 1). Furthermore, the QnR map can be nonlinear and especially in the case of pronounced secondary QnR maxima (marked by black squares in iii & iv) user intervention is required to judge which parameter set yields the best compromise of resolution and fidelity. Scale bar in a&b 5 µm, inset i-iv) scale bars 2 µm.