Quantification of astrocytic synaptic pruning in hippocampus in response to in vitro Aβ oligomer incubation via colocalization analysis with C1q

Quantification of synaptic engulfment is an indirect measurement of synaptic pruning. Here, we provide a detailed protocol for the volumetric rendering of individual high-resolution astrocytes in the CA1 region of hippocampus in an in vitro slice model of Amyloid-beta (Aβ) treatment. This includes free floating slice preparation, treatment with Aβ oligomers, immunofluorescence, confocal imaging and analysis of individual astrocytes. We also provide a comprehensive analysis for 3D rendering of astrocytes and assessment of synaptic engulfment via “eat-me tag” C1q protein and synaptic marker PSD95. Highlights ![Figure][1] ### Competing Interest Statement The authors have declared no competing interest. [1]: pending:yes