Figure 2.

Schematic figure depicting current methods adopted to obtain freshly isolated microglia. Brains from adult mice are (1) homogenized with a dounce homogenizer at 4°C to avoid microglia activation or (2) enzymatically digested at 37°C in the water bath, and filtered through a 70 μm cell strainer. Myelin is removed either by centrifugation with a Percoll gradient or by using anti-myelin magnetic cell sorting (MACS) antibodies. Then, microglia are sorted from other CNS cell types by FACS through specific cell markers. Otherwise, microglia are isolated by MACS anti-CD11b antibodies. Microglia can also be isolated by immunopanning. In this approach, the cells suspension is passed over a series of antibody coated dishes in order to remove contaminating cell types. Then, microglia are positively isolated in the last coated dish.