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Primary neuron culture for live imaging of axonal cargoes
Output Details
Description
This protocol describes the preparation and culture of mouse primary cortical neurons for live-imaging experiments. Cortices were dissected from mouse embryos at day 15.5. Cortical neurons were isolated by digestion with 0.25% Trypsin and trituration with a serological pipette. Neurons were plated on glass-bottom imaging dishes in Attachment Media. After 5 hours in culture, Attachment Media was replaced with Maintenance Media, and AraC was added on the next day to prevent glia cell proliferation. Neurons were transfected 16-24 hours before imaging using Lipofectamine 2000.