Fig. 5: ExPRESSO enables enhanced interrogation of cells and non-nuclear features, including the BBB, in archival human brain sections.

a MIBI imaging of an ExPRESSO-processed human hippocampus tissue section (top) and middle frontal gyrus (bottom) with tile acquisition of 9 FOVs, each one with a size of 400x400 μm (physical measurements). Shown are markers for pre-synapse (Synaptophysin), astrocyte projections (GFAP), nuclei (Histone H3), microglia (Iba1), vessel lumen and BBB leakiness (Fibrinogen or Albumin or SAA and Fibrinogen), vessels (GLUT1), muscularized vasculature (SMA), basement membrane (Collagen IV), endothelial cells (CD105 and vWF), astrocytes (GlnSyn or GFAP), neurons (MAP2), pre-synapse (VAMP2 or Synaptophysin), neurons with oxidative stress (8OHG), myelin and oligodendrocytes (MAG). Enlarged views of boxed areas are shown as insets. Scale bars (the ExPRESSO-MIBI scale bars indicate the pre-expansion dimensions based on a 3.7-fold expansion): 100 μm (main) and 20 μm (enlarged). b Anchoring analysis around vessel feature segment components on both MIBI samples (left) and ExPRESSO-MIBI gels (right) in both curve and heatmap. The average counts were plotted as solid lines, while the 95% confidence intervals were plotted as shadow-around lines. The heatmap of ExPRESSO-MIBI is labeled with blue bars to indicate the local maximum of the respective channel in the ExPRESSO gel analysis. c Left: Schematic of components of the BBB and markers used for their identification in the ExPRESSO-MIBI brain panel. Middle and right: Representative complex of BBB, enlarged view from a region in Fig. 5a, and bicolor enlarged regions to further highlight imaging details. Scale bars (the ExPRESSO-MIBI scale bars indicate the pre-expansion dimensions based on a 3.7-fold expansion): 250 μm (middle) and 10 μm (right, enlarged).