Dual AO/EB staining to detect apoptosis in osteosarcoma cells compared with flow cytometry

Area of Science:

• Cell biology
• Cancer research
• Biotechnology

Background:

• Apoptosis involves distinct cell membrane changes.
• Differentiating normal, apoptotic, and necrotic cells is crucial in cancer studies.
• Acridine orange/ethidium bromide (AO/EB) staining visualizes these cellular changes.

Purpose of the Study:

• To evaluate dual acridine orange/ethidium bromide (AO/EB) staining for detecting tumor cell apoptosis.
• To compare AO/EB staining with flow cytometry for apoptosis assessment.

Main Methods:

• Human osteosarcoma cells were cultured with varying concentrations of kappa-selenocarrageenan.
• Cells were stained with acridine orange/ethidium bromide (AO/EB) or propidium iodide (PI).
• Apoptosis rates were analyzed using fluorescent microscopy and flow cytometry.

Main Results:

• Distinct fluorescent patterns identified normal, early apoptotic, late apoptotic, and necrotic cells.
• Early apoptotic cells showed yellow-green nuclear staining.
• Late apoptotic cells exhibited concentrated orange nuclear staining, while necrotic cells displayed peripheral orange-red fluorescence.
• AO/EB staining results for apoptosis rates were comparable to flow cytometry (P>0.05).

Conclusions:

• Dual acridine orange/ethidium bromide (AO/EB) staining is a viable method for detecting tumor cell apoptosis.
• AO/EB staining offers an economical and convenient approach for evaluating tumor chemosensitivity.