Summary
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
Measurement of synaptic engulfment in response to treatment with Aβ peptide
Volumetric reconstruction of high resolution individual astrocyte
Colocalization analysis of astrocyte and complementary “eat-me” protein C1q
Competing Interest Statement
The authors have declared no competing interest.