Fig 1: Immunofluorescence analysis of fiber-high HSCR tissue for neuronal marker. (A) Immunofluorescence (5-µm Swiss roll cryosections) of aganglionic rectum region from a fiber-high patient using tubulin (Alexa647, green), a general neuronal marker, combined with VIP (A555, red), labeling peptidergic neurons; TH (A555, red), labeling dopaminergic and adrenergic neurons; NOS (A555, red), labeling nitrinergic neurons, and S100B (S100 calcium-binding protein B; A555, red), labeling glia cells. DAPI (blue) shows cell nuclei. Images were taken by fluorescence microscopy (original magnification, ×20; scale bar, 100 µm) and processed by Fiji software. Close-up views (white boxes) show mucosal/epithelial region. Negative control: immunofluorescence (5-µm Swiss roll cryosections) of aganglionic rectum region from a fiber-high patient using secondary antibody controls. (B) Immunofluorescence of myenteric plexus ganglia from ganglionic control tissue using the antibody combinations from (A). Scale bar, 50 µm; scale bar for NOS images, 30 µm.