7AAD Staining of Nonviable Cells

1. Add 1µl of 1 mg/ml 7-AAD (1µg/ml final) to approximately 1x10⁶ washed cells suspended in 1ml PBS.
2. Incubate ≥ 30 minutes in the dark on ice.
3. Analyze on flow cytometer with excitation at 488nm. Collect fluorescence emission with a 650nm long-pass or a 670±20nm band-pass filter.

7-AAD is easily excitable at 488nm. The fluorescence emission of the dye has a peak at ~670nm.

From Current Protocols in Flow Cytometry