Microglia were isolated as previously described [11 (link)]. Mice were anesthetized and hippocampi were dissociated from the brain and transferred to HBSS buffer. A neural dissociation kit (product No. 130-092-628, Miltenyi Biotec, Waltham, MA) was used to prepare a single-cell suspension, followed by a continuous 30% Percoll gradient at 700 g for 15 min. For microglia/macrophage isolation, cells were incubated with biotin-conjugated anti-CD11b antibody (eBioscience product no. 13-0112-82, Thermo Fisher Scientific, Waltham, MA), and CD11b + cells were isolated using Dynabeads Biotin Binder (Invitrogen product no. 11047, Thermo Fisher Scientific) and following manufacturer’s instruction.
Protocol detail
Isolation of Microglial Cells from Mouse Hippocampus
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Other organizations : Yale University, Columbia University
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Variable analysis
independent variables
- Microglia isolation method (using CD11b antibody and Dynabeads)
- Not explicitly mentioned
- Anesthesia of mice
- Dissociation of hippocampi from the brain
- Preparation of single-cell suspension using neural dissociation kit
- Continuous 30% Percoll gradient centrifugation
- Positive control: Not mentioned
- Negative control: Not mentioned
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