AO/PI (acridine orange / propidium iodide) Staining Solution for live/dead Mammalian nucleated cells
The ViaStain™ AO/PI Staining Solution in PBS enables the user to quantitatively distinguish live and dead nucleated cells from a variety of primary mammalian cell samples, even in the presence of a high background of non-lysed red blood cells, platelets and/or debris using the Cellometer system. This formulation has been optimized to work with whole blood, peripheral blood mononuclear cells (PBMC), bone marrow, splenocytes, thymocytes, lymph nodes and hepatocytes but also works in various other digested tissues and cultured cell lines. The solution contains a combination of the green-fluorescent nucleic acid stain, acridine orange, and the red-fluorescent nucleic acid stain, propidium iodide. Propidium iodide is a membrane exclusion dye that only enters cells with compromised membranes while acridine orange penetrates all cells in a population. When both dyes are present in the nucleus, propidium iodide causes a reduction in acridine orange fluorescence by fluorescence resonance energy transfer (FRET). As a result, nucleated cells with intact membranes stain fluorescent green and are counted as live, whereas nucleated cells with compromised membranes only stain fluorescent red and are counted as dead when using the Cellometer system. Non-nucleated material such as red blood cells, platelets and debris do not fluoresce and are ignored by the Cellometer software.