Perinuclear VIFs are loosely aligned within disorganized bundles. (A) 3D bounding box corresponding to the analyzed perinuclear FIB-SEM volume. (B) Representative slice cropped from this volume depicting staining density within the crowded cytoplasm. Scale bar = 200 nm. (C) Cropped FIB-SEM slice with magnified insets indicating cross-sectional profiles of a VIF (top), an MT (center), and a ribosome (bottom); different z slices from this stack are represented in Fig. 8 G. (D) Orthogonal FIB-SEM slices centered on a VIF bundle indicating representative transverse and longitudinal VIF profiles. (E–G) 3D reconstructions of VIFs (blue) traced from the FIB-SEM volume. The ventral plasma membrane is indicated in gray. (H) Maximum intensity projection of a FIB-SEM subvolume featuring a thick vimentin bundle (blue overlay). Scale = 200 nm. (I) Segmented filaments (blue dots) in a cross section through a VIF bundle. The four shaded areas indicate subregions of filament clustering within the larger bundle architecture. (J) 3D rendering of a VIF bundle clipped at three positions to reveal differences in relative filament positioning. (K) Aligned montage of isolated VIF bundles derived from the volume indicated in A. (L) Scatter plot indicating average orientation relative to first nearest neighbors and average distance to first nearest neighbors for traced filaments. The dashed line indicates 20° threshold for co-orientation. (M) Rendering of perinuclear VIFs color-coded by orientation relative to nearest neighboring filaments. Blue indicates aligned filaments (relative orientation <20°), while gray indicates nonaligned filaments (relative orientation >20°). (N) Magnified inset indicating blue aligned filaments within a bundle and neighboring gray, nonaligned, curved VIFs.