Bacteria Viability Protocols

Propidium Iodide (PI)

1. Harvest bacteria
2. Add 5 mg/ml PI to 1ml of bacteria
3. Incubate at 37C for 10 minutes
4. Analyse 50,000 events by flow cytometry
5. Change FSC and SSC to Log scale and adjust Threshold to 200 on BD LSRII
6. Excite at 488nm wavelengths collect emission at 610 nm

DAPI

1. Harvest bacteria
2. Add 200ng DAPI to 1ml of bacteria
3. Incubate for 10 minutes at Room Temperature
4. Analyse 50,000 events by flow cytometry
5. Change FSC and SSC to Log scale and adjust Threshold to 200 on BD LSRII
6. Excite with UV or 405nm laser collect emission at 461nm