Fig. 1

Column-free immunomagnetic separation generates highly pure cultures of microglia. Mixed glia (a–c), microglial (d–f), and astrocyte (g–i) cultures were assessed for total glia composition via immunofluorescence for GFAP-positive (red) and IBA-1-positive (green) cells or via flow cytometry for Cd11b and GLAST. Representative ×20 images of mixed glia (a), microglial (b), and astrocyte (c) cultures with a ×40 insert showing GFAP (red), IBA-1 (green), and DAPI (blue). Scale bars = 10 μm. Quantitative counts were determined for the number of glia present in mixed glia (b), microglia (e), and astrocyte (h) cultures both by positive immunoreactivity (colored) or by consistent morphology in the absence of positive staining (gray). Data are presented as mean percent of total cells per field ± SEM. Flow cytometry scatter plots showing the percentage of Cd11b- or GLAST-positive cells for mixed glia (c), microglia (f), or astrocyte (i) cultures