Extended Data Fig. 12: Related to Fig. 4h–j and spatially restricted variation in helical twist and polarity orientation of in-tissue tau filament clusters.

a, Left and right, resolution estimation of subtomogram averaging of 52 PHF and 19 SF in a single tissue cryoFIB-SEM lift-out lamella tomogram by gold-standard Fourier shell correlation (FSC), respectively. b, Resolution estimation of subtomogram averaging of 64 tau SF in a single tissue cryoFIB-SEM lift-out lamella tomogram by Fourier shell correlation (FSC). c, Filament helical twist (left y-axis) and cross-over distance (right y-axis) of in-tissue tau filament clusters in different locations of AD post-mortem brain from seven cryo-section (CS1-7) and two lift-out lamella tomograms, one of which (LOL1) was composed of two ultrastructural polymorphs, related to maps shown in Fig. 3g, Fig. 4g & i and Extended Data Fig. 9a–f. Orange and cyan data points, tau cluster subtomogram averages that fitted the atomic models of PHF or SF using EM placement (see Supplementary Data Table 5), respectively. Yellow data points, tau cluster subtomogram averages in which the ultrastructural polymorph was unresolved. For reference, the helical twist/crossover distances calculated for AD PHF and SF are shown in grey (data point and whiskers, cryoEM and range of twists observed by negative stain EM from Fitzpatrick et al.²⁴, respectively). See also Supplementary Data Table 5. d, Filament subtomogram average map (see Fig. 3h) mapped back into the raw tomographic volume with PHF filaments coloured by polarity orientation. Orange and indigo filaments with upwards and downwards polarity orientation, respectively. Top and bottom, volume viewed from top and rotated ~45°, with and without tomographic slice, respectively. Scale bar, 10 nm. e, Same as b, but related to Fig. 4h. Cyan and purple, SF filaments with upwards and downwards polarity orientation, respectively. f, Same as b and c, but related to Fig. 4j.