Flow chart of CD11b
cell isolation. Animals were perfused with ice-cold PBS to remove circulating immune cells. Brains were dissected and tissue was gently dissociated using enzymatic digestion. Myelin was removed using one of three different methods (Percoll or sucrose centrifugation, or myelin magnetic beads). Cells were stained with anti-CD11b antibodies and then incubated with magnetic beads. Samples were extensively washed on columns in a magnetic field to obtain the CD11b+ or effluent (CD11b-) cell fractions. The total time for isolation was less than 3 hours.