Quantifying IBA1 Expression in SCI

We used NanoZoomer RS slide scanner with constant light intensity and exposure time and NanoZoomer Digital Pathology System view software (Hamamatsu, Hamamatsu, Japan). To quantify SCI and GW2580 treatment-induced changes in IBA1 expression, the mean optical density (OD) was measured along the spinal cord, as previously described (ImageJ, National Institutes of Health, USA) 23 (link), 33 (link), 34 (link), 37 . To minimize bias in staining intensity, all immunostainings for a given antibody and a given time-point were done in parallel. For all antibodies used, expression levels were analyzed in at least 40 and 16 axial sections throughout the lesion segment of the spinal cord at 210 µm and 630 µm intervals for Microcebus Murinus and mice respectively. OD quantifications included grey and white matters (excluding the dorsal funiculus) and dorsal funiculus. Background was subtracted from OD values of each section. All quantifications were done blindly.