Possible causes for this rapid pH change can be:
– Inappropriate carbon dioxide pressure: Increasing or decreasing the percentage of carbon dioxide in the incubator is based on the concentration of sodium bicarbonate in the medium. For concentrations of 2 to 3.7 g/L sodium bicarbonate set the 5-10% carbon dioxide.
– Tightly close the cap of cell culture flasks then screw a quarter.
– Insufficient bicarbonate buffer: Add HEPES buffer to the final concentration of 10-25 mM.
– Bacterial, fungal or yeast contamination
HEPES is widely used in cell culture as a buffering reagent to maintain physiological pH. However, some protocols recommend without HEPES cell culture media.
For more information about the exact amount, you can refer to the datasheet of each product.
The amount of glucose or other carbohydrates is specific to each product. Please refer to the datasheet of each medium.
Phenol red is a water-soluble coloring reagent that is used in culture media as a pH indicator from yellow to red “ph 6.6 to ph 8” and bright pink for pH above 8.1. Phenol red is widely used in cultured media formulations to provide a visual indicator of pH changes.
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