After cultured primary forebrain neurons to DIV15, media was removed from the wells and propidium iodide (PI) dissolved in HBSS was added at 100 μM. The cells were incubated for 2 min followed by the addition of equivalent volume of HBSS containing the cell-permeable fluorescein diacetate (FDA) yielding final concentrations of 12 μM for FDA and 50 μM for PI. Cells were imaged 5 min later.

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