Remove the culture media from the wells, and immediately treat the cells with 150 microliters of a test formulation or media control solution. Then, incubate the cells for 30 minutes.
Remove the test solution from the cells, and add 0.5 milliliters of 10% metabolic dye solution. Incubate the cells for another four hours.
After the incubation, remove 100 microliters of dye solution from each well and transfer it to a 96-well plate. Use a plate reader to measure the fluorescence of each well, setting the excitation to 540 nanometers and emission to 590 nanometers.
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