Colorimetric Method for Estimating Number of Cells in Monolayer Cultures without Physiological Damage.

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Abstract
A method is presented for determining the number of cells in monolayer cultures of monkey kidney cells without physiologically damaging them. The method consists of determining the change in absorption of a phenol red solution after a 15-second exposure of the cultures under standard conditions. These conditions include the use of a standard solution defined with respect to pH, quantity of titratable phenol red, and buffering salt, and the use of the same type of glass in the different cultures. The change in absorption appears to be a simple consequence of the production of hydrogen ion by the cells. It is critically dependent on the presence of potassium but independent of an exogenous carbon source, and, independent of endogenous polysaccharide reserves over a wide range. The amount of hydrogen ion product may be sufficiently independent of cell age and metabolism to be a reliable method for the estimation of growth in cultures.