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Application Note

Comparison of trypan blue vs acridine orange/propidium iodide staining methods

Cell viability assessment is essential across biological experiments, from routine cell culture to drug testing on tumor cells. Trypan blue (TB) exclusion has been a standard method for decades1,2, but studies over the past 20 years show TB consistently reports higher viability than fluorescence-based assays in time-course measurements1, 3-5.

While these differences are documented, the underlying reasons remain unclear. We conducted a time-course study comparing TB with acridine orange/propidium iodide (AO/PI) using automated image-based cytometry to investigate these discrepancies.

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Comparison of trypan blue vs acridine orange/propidium iodide staining methods

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