Figure 3

Histopathological analysis of the non-human primate brain tissue following focused ultrasound treatment. (a) Gross pathology and corresponding BBB opening area at the 2-day (left) and 18-day (right) time point. (b) H&E staining and corresponding BBB opening area at the 2-day (left) and 18-day (right) time point. (c) Luxol fast blue H&E staining for myelin delineation at the 2-day (i–iv) and 18-day (v–viii) time point. (d) GFAP staining for astrocyte presence at the 2-day (i–iv) and 18-day (v–viii) time point. (e) Iba1/CD68 staining for microglia presence and migration at the 2-day (i–iv) and 18-day (v–viii) time point. (f) DCX staining for immature neurons at the 2-day (i–iv) and 18-day (v–viii) time point. GFAP and DAPI were used to stain astrocytes and cell nuclei, respectively. (g) Iba1⁺ and CD68⁺ cell density within perivascular areas treated at MI of 0.4 and 0.8, examined at the 2-day (deep blue bars) and 18-day (purple bars) time points. (h) DCX⁺ cell density with brain regions treated at MI of 0.4 and 0.8, examined at the 2-day (light blue bars) and 18-day (pink bars) time points. Regions of interest at (c), (d), (e), and (f) were within the left hemisphere treated at MI of 0.4 (i, ii, v, vi) or within the right hemisphere treated at MI of 0.8 (iii, iv, vii, viii). Color bars: normalized contrast enhancement. Scale bars: 1 cm (a, b); 1 mm (c); 200 μm (d, e); 100 μm (d-iv, e-iii, f). All images shown here correspond to regions of interest within areas with BBB opening as confirmed by MRI. Green shaded areas in (g) and (f) correspond to baseline Iba1⁺-CD68⁺ and DCX⁺ cell density, in brain areas outside the treated volume. Data in (g) and (h) are presented as mean ± standard deviation (n = 5 regions of interest per section and per hemisphere, n = 2 sections). GP, gross pathology; H&E, hematoxylin and eosin; CE-T₁, contrast-enhanced T₁ MRI scan; LFB, luxol fast blue for myelin delineation; GFAP, glial fibrillary acidic protein; Iba1, ionized calcium binding adaptor molecule 1; CD68, cluster of differentiation 68; DCX, doublecortin; DAPI, 4′,6-diamidino-2-phenylindole.