Figure 2 Histochemical and immunohistochemical staining and electron microscopic examination of structures in the brain biopsy Hematoxylin & eosin staining.

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Figure 2 Histochemical and immunohistochemical staining and electron microscopic examination of structures in the brain biopsy Hematoxylin & eosin staining (A), periodic acid-Schiff (PAS) staining (B), and Grocott staining (C) of brain biopsy samples show the presence of round pathogens, similar to Michaelis-Gutmann bodies, located mainly around the blood vessels. Histochemical and immunohistochemical staining and electron microscopic examination of structures in the brain biopsy Hematoxylin & eosin staining (A), periodic acid-Schiff (PAS) staining (B), and Grocott staining (C) of brain biopsy samples show the presence of round pathogens, similar to Michaelis-Gutmann bodies, located mainly around the blood vessels. Some CD4-positive T lymphocytes are also located around the vessels (D). CD8-positive T lymphocytes appear to be the main invasive cells (E), whereas CD68-positive macrophages are only present in patches (F). Toluidine blue and Safranin O stain of Epon-embedded semithin sections show some pathogens without nuclei around vessels (G). Electron microscopy (H–J) shows an irregular shape of pathogens (diameter 2–7 μm) without cell walls and nuclei (arrows) but with some membranous structures in the cytoplasm (triangles). Scale bar = 5 μm. Brain biopsies from patients 2 (K), 3 (L), and 4 (M) demonstrate a similar histopathology to that of patient 1, with 2–7 μm round PAS-positive pathogens located mainly around the vessels. The agents showed an irregular shape with no detectable cell wall or nucleus under an electron microscope (arrows). Scale bar = 5 μm. Yusuke Sakiyama et al. Neurol Neuroimmunol Neuroinflamm 2015;2:e143 © 2015 American Academy of Neurology