glutamate receptor AMPA was investigated and CNQX, a commonly used competitive AMPA receptor antagonist, was used to silence activity due to glutamatergic inputs from excitatory neurons in the culture. CNQX was added to the cultures during experiments at a final concentration of 20 μM. Images were collected at approximately 9 Hz. Reagents were dispensed at frame 900 for CNO or clozapine treatment or at frame 1200 for CNQX addition. After an additional 1500–1800 frames were collected, the analysis was complete, and the reagent plate was switched out for a plate containing High KCl buffer. An additional 600 or 900 frames were collected after the plates were exchanged, with a transfer of 80 μl KCl occurring at frame 300 of the additional set. The KCl addition depolarized all neuronal cells and verified cell viability at the end of each experiment.
