alamarBlue Assay: A Sensitive Fluorometric Method to Screen the Cytotoxic Effects of Artificial Tear Formulations on Metabolic Activity of Human Corneal Epithelial Cells

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.