Fig. 1. Identification of microfeatures using immunohistochemistry and corresponding nanostructure characteristics. (A) LBs and SOD1 aggregates in 20 μm fresh frozen SN sections are immunopositive for anti-human phosphorylated (pS129) α-synuclein, and anti-human misfolded B8H10 SOD1, respectively using immunofluorescence, while NM can be visualized in the native state using brightfield microscopy. Feature coordinates were used to identify corresponding regions of interest in facing sections mounted on Si3N4 windows. Simultaneous X-ray ptychography (B) and XFM (C) was used to construct spatial phase contrast (darker areas represent greater compositional diversity) and electron density (as Compton scatter, with lighter areas representing higher density) images of each identified feature. (D and E) Skewed histograms of phase contrast images reflect compositional diversity in the depth of structures, while normal frequency distributions from Compton scatter maps depict photon noise only. Scale bars = 1 μm. Representative images from scan IDs fly6 (LB), fly96 (SOD1), and fly67 (NM; see ESI Fig. 1† for corresponding elemental maps).
