(A) Simulated widefield impβ localization at the equatorial plane of the nucleus and (B) corresponding dSTORM image. Mean localization precision is 12 nm. (C and D) Corresponding widefield and dSTORM images taken at the basal surface of the nucleus showing characteristic punctate NPC structures. (E) Zoom of the dSTORM image in (B), showing discrete NPC structures (examples, blue arrows). Putative NPC structures (green) markers are automatically identified using the linearized envelope localization histogram (shown in white, Supplementary methods). (F) Method for putative NPC structure isolation and alignment. Peaks in the envelope histogram (black line) are identified as potential locations for putative NPC structures (black circles). Localizations falling into a window (width, w, length, l) centred at these locations (p) are cropped out and rotated to a common frame, c, by the angle θ, maintaining the cytoplasm-nucleus vector, n.