Hello, I have carried out similar experiments but depleted microglia and experimented on astrocytes. I found that 48 hr after isolation was sufficient. I would advise assessing morphology under a light microscope to give you an idea of when is best to start your experiment.

Hey Deborah,

I also isolated microglia from adult brain using the miltenyi kits and MACS and I tried several time points (24h 48h and 72h). I experienced no differences between 24h and 48h but the expression of some marker genes changes after 72h. Based on this, I let the cells recover for 24h before using for further experiments.

All the best and good luck with your experiments.