Fig. 1

Double immunofluorescence labeling of cerebellar frozen sections from a human autoptic case. Binding sites for autoantibodies from the patient (green) and astroglial or neuronal markers (red) are concomitantly revealed by confocal laser-scanning microscopy. Autoantigens are stained by patient’s serum (1:20) and carbocyanine (Cy) 2-conjugated anti-human IgG + IgM, while astroglia and neurons are visualized by appropriate Cy3-coupled secondary antibodies. a At lower magnification, the cerebellar subcortex displays layers of granular cells immunoreactive for the patient’s serum—clearly distinguishable from immunoreactivity for rabbit-anti-glial fibrillary acidic protein (GFAP; Dakocytomation; 1:1000). b At higher magnification, autoantibodies bind cells of deep cerebellar nuclei lacking GFAP immunolabeling. c In the cerebellar cortex, Purkinje cells are stained by patient’s serum while rabbit-anti-S100β (Swant; 1:500) predominantly demonstrates protoplasmic astroglia. d Immunodecoration of probably neuronal surface antigens with patient’s serum is located apart from labeling achieved with mouse-anti-neuronal nuclei (NeuN; Millipore; 1:100) in the layer of deep cerebellar neurons. Scale bars in a, c = 200 µm, in b, d = 50 µm