Figure 3

Double fluorescence immunohistochemistry of oligodendrocytes, astrocytes and microglia. Double fluorescence immunostaining for O4 and CD59 or C1-inh is demonstrated in A-F. In A and D, cells of typical oligodendroglial morphology were stained in the initial cycle for the specific oligodendroglial marker O4. Detection is by a Texas Red-conjugated secondary antibody. Second cycle staining for CD59 (B) and C1-inh (E) are shown. The detections are by an FITC-linked green fluorescent secondary antibody. In C and F, double immunofluorescences are shown in which the cells appear yellow, demonstrating colocalization of O4 with CD59 or C1-inh. Double fluorescence immunostaining of astrocytes for GFAP and CD59 or C1-inh is demonstrated in G-L. In G and J, cells of typical astrocytic morphology are stained in the initial cycle for the specific astroglial marker GFAP. Detection is by a Texas Red-conjugated secondary antibody. Second cycle staining for CD59 (H) and C1-inh (K) is shown with an FITC-linked green fluorescent secondary antibody. In I and L, double immunofluorescences are shown in which the cells appear yellow, demonstrating colocalization of GFAP with CD59 or C1-inh. Double fluorescence immunostaining for microglia using the specific marker CD68 and CD59 or C1-inh is demonstrated in M-R. In M and P, cells of typical microglial morphology are stained by CD68 with detection by a Texas Red-conjugated secondary antibody. Second cycle staining for CD59 (N) and C1-inh (Q) are shown. The detections are by an FITC-linked green fluorescent secondary antibody. In O and R, double immunofluorescences are shown in which the cells appear yellow, demonstrating colocalization of CD68 with CD59 or C1-inh. (Magnification: × 200)