Figure 1.
3D single-molecule localization microscopy (SMLM)of clathrin-coated structures. (A) 3D SMLM image of clathrin immunolabeled with AF647 at the bottom membrane of a SK-MEL-2 cell. (B) Enlarged view of the region indicated in A. (C) All structures found in B are shown in top view (xy) and as 50 nm-thick z-slices (xz), and orientation of slice is indicated by a dotted line. Scale bars are 10 µm (A); 1 µm (B); and 100 nm (C). (D) Geometric analysis pipeline. All clathrin coats are segmented, and their localizations are directly fitted with a spherical cap model using the fitting framework LocMoFit (Wu et al., 2023). (E) In LocMoFit, individual clathrin coats are parametrized by their size (radius R), closing angle (θ), position (x0,y0,z0), and 3D orientation. (F) Using θ as a proxy for endocytic progression, the relative endocytic time point for each structure can be determined.

3D single-molecule localization microscopy (SMLM) of clathrin-coated structures. (A) 3D SMLM image of clathrin immunolabeled with AF647 at the bottom membrane of a SK-MEL-2 cell. (B) Enlarged view of the region indicated in A. (C) All structures found in B are shown in top view (xy) and as 50 nm-thick z-slices (xz), and orientation of slice is indicated by a dotted line. Scale bars are 10 µm (A); 1 µm (B); and 100 nm (C). (D) Geometric analysis pipeline. All clathrin coats are segmented, and their localizations are directly fitted with a spherical cap model using the fitting framework LocMoFit (Wu et al., 2023). (E) In LocMoFit, individual clathrin coats are parametrized by their size (radius R), closing angle (θ), position (x0,y0,z0), and 3D orientation. (F) Using θ as a proxy for endocytic progression, the relative endocytic time point for each structure can be determined.

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