JoVE Encyclopedia of Experiments
Immunology
0 views • 4:45 min • July 8th, 2025
Microglia — the brain-resident macrophages — express CD11b, a cell-surface integrin.
To isolate microglia from mouse pup brains, take cerebrum fragments in dissociation tubes containing buffer with papain, a proteolytic enzyme, and deoxyribonuclease.
Place the tubes on a mechanical dissociator. During the run, the dissociator mechanically disrupts the tissue.
Papain digests the tissue’s extracellular matrix, releasing cells, including microglia, into suspension. Deoxyribonuclease degrades free DNA in suspension, preventing inefficient cell isolation.
Centrifuge. Complete the mechanical dissociation by pipetting. Transfer the suspension through a cell strainer to remove debris and cell clumps, and obtain a homogeneous single-cell population.
Incubate with superparamagnetic microbeads functionalized with anti-CD11b antibodies. The microbeads specifically bind to CD11b on cells, including microglia.
Post-incubation, centrifuge. Remove the supernatant containing unbound beads. Resuspend the cells in buffer. Load onto a column placed on a magnetic separator. The column comprises a matrix with ferromagnetic beads.
When placed on the separator, the spheres within the column amplify the ma
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