Figure 4
Surgical histopathology of Patient 9 (table 1). (A) Concentration of dysmorphic neurons (arrow) decorated with anti-non-phosphorylated neurofilament H-specific antibodies (clone SMI32, Alexa555-labeled anti-mouse IgG1 secondary antibody, orange pseudocolour) at the bottom of a sulcus (sulcal surface indicated by small arrowheads), with concomitant accumulation of vimentin-positive balloon cells (clone SP20, Alexa488-labeled anti-rabbit IgG secondary antibody, green pseudocolour; arrowheads) in the underlying white matter. In addition, vascular myocytes expressing smooth muscle actin (clone 1A4, Alexa647-labelled anti-murine IgG2a secondary antibody, magenta pseudocolour) and nuclei (Hoechst 33342, blue pseudocolour) are visualized (multichannel-immunofluorescence whole slide imaging, 3DHistech MIDI). (B) High-power magnification of area indicated by asterisk in (A) showing predominant localization of vimentin-immunopositive balloon cells (BC) in white matter (as indicated by arrows) and SMI32-immunopositive dysmorphic neurons (DN) in grey matter. Scale bar in (A) = 2 mm, in (B) = 50 μm.