Extended Data Fig. 8: Validation of in situ PSF modelling for imaging a thick sample.

Imaging of Nup96-SNAP-AF647 in U2OS cells was performed using an oil immersion objective lens (NA = 1.5) and imaging buffer (n = 1.406, Methods) on a single-channel SMLM system equipped with a deformable mirror (DM). (a) A sandwiched sample of U2OS cells was prepared for an imaging depth of 25 µm. (b) Top view of the reconstructed Nup96 using the in situ PSF model. (c) Enlarged XY view of the selected region in (b). (d) XZ view of the selected region in (c) reconstructed from bead and in situ PSF models, respectively. Scale bars, 2 µm (b), 1 µm (c), and 0.5 µm (d).