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Ethidium Bromide/Acridine Orange

 

For discrimination of live from dead cells on the basis of membrane integrity.

•Acridine orange fluoresces GREEN --> LIVE cells.

•Ethidium bromide fluoresces orange (intercalates with DNA) --> DEAD cells.

 

From powder:

  • 15 mg acridine orange
  • 50 mg ethidium bromide dissolved in 1 ml 95% ethanol
  • add 49 ml H 2 0, mix well, freeze in 1 ml aliquots.

 

Working conc for cell viabilities:

Take 1 ml & make up to 100 ml in PBS

 

From solutions:

  • 300µL of 10 mg/mL acridine orange
  • 1000µL of 10 mg/mL ethidium bromide
  • Make up to 100mL with PBS