Bacteria Membrane Potential Protocols

Bis-oxonal DiBAC4(5) with DAPI

1. Harvest bacteria
2. Add 10mg/ml of Bis-oxonal dye to 1ml 0.55 OD of bacteria
3. Incubate for 10 minutes at Room Temperature
4. Wash twice in PBS
5. Add 200ng/ml DAPI
6. Analyse 50,000 events by flow cytometry
7. Change FSC and SSC to Log scale and adjust Threshold to 200 on BD LSRII
8. Excite with 488nm laser collect emission at 575nm