Coat a 24-well culture dish with 0.01% Poly-L-lysine overnight at 37 °C. Wash three times with ddH₂O the following day.
Prepare the following cell culture medium: DMEM containing 10% FBS, 1% penicillin/streptomycin, and 2 mM L-glutamine.
(Optional) Add 20 ng/mL M-CSF (#130-094-129) to enhance microglia growth and survival.
Use the Neural Tissue Dissociation Kit (T) or the Neural Tissue Dissociation Kit (P) to dissociate brain tissue and prepare a single-cell suspension. Follow the protocol of the kit data sheet.
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Isolate microglia from the single-cell suspension using either the CD11b (Microglia) MicroBeads, human and mouse or the CD11b/c (Microglia) MicroBeads, rat. Follow the protocol of the corresponding kit data sheet.
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CD11b+ microglia isolated from neonatal mouse brain tissue. CD11b+ cells were isolated from mouse neural cell suspension using the CD11b (Microglia) MicroBeads, an MS Column, and a MiniMACS™ Separator. Cells were fluorescently stained with CD11b-APC and CD45-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.
CD11b/c+ microglia isolated from neonatal rat brain tissue. CD11b/c+ cells were isolated from P3 rat brain tissue using the Neural Tissue Dissociation Kit (P), the gentleMACS™ Dissociator, CD11b/c (Microglia) MicroBeads, a MiniMACS™ Separator, and two MS Columns. Cells were fluorecently stained with CD11b/c-FITC in combination with CD45-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.
|Column||Max. number of labeled cells||Max. number of total cells||Separator|
|Positive selection or depletion|
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