Cy3 conjugated primary antibody against gfap

To determine the extent of the glial scar, immunohistochemical analysis of a CY3-conjugated primary antibody against GFAP (Sigma, St. Louis, MO, USA) was used. Samples from five animals from each group were obtained. Each sample of the spinal cord was cut at 1 mm intervals. A total number of 15 cross sections, including the center of the lesion, was observed with an Axioskop 2 plus microscope (Zeiss, Oberkochen, Germany). The acquired images were analyzed using ImageJ software (NIH, Bethesda, MD, USA). The GFAP positive area around the central cavity (Figure 4(B1)) together with the number of protoplasmic astrocytes (Figure 4(B2,B3)) was measured on each section.

Immunohistochemical visualization of the astrogliosis and protoplasmic astrocytes was obtained by staining with CY3-conjugated primary antibody against GFAP (1:400, Sigma-Aldrich, St. Louis, MO, USA) (Figure 4A,B). From each group, five animals were obtained. A total number of ten cross-sections, including the center of the lesion and areas both cranially and caudally, were observed and photographed with an Axioskop 2 plus microscope (Zeiss, Oberkochen, Germany). Acquired images were evaluated for the total area of GFAP high intensity signal around the malatic cavity representing the astrogliosis by ImageJ software (NIH, Bethesda, MD, USA). On the same images, the total number of protoplasmic astrocytes was manually counted (Figure 4C).