Protocol to assess engulfment and degradation of synaptosomes by murine microglia in vitro.

Microglia, the innate immune cells of the central nervous system, refine neuronal circuitries both during brain development and in neurodegenerative diseases. Here, we present a protocol to independently assess the engulfment and degradation of synaptosomes by murine microglia, both in fixed and live samples, using confocal imaging. We describe steps for isolating primary microglia, preparing and conjugating synaptosomes with pHrodo, and performing an uptake and degradation assay. We then detail procedures for analyzing synaptosome uptake and degradation using ImageJ software. For complete details on the use and execution of this protocol, please refer to Matera, Compagnion, et al.(1).