AAT Bioquest

How do I measure cell proliferation?

Posted August 23, 2022


There are 4 techniques you can use to measure cell proliferation. The method you choose should depend on the type of cell you are studying, your research protocols, and the type of information you are looking to gain from the cell proliferation assay.

Using DNA synthesis assays

This is the most reliable and accurate way to measure cell proliferation. It involves incubating radiolabeled 3H-thymine with cells for several hours. Cell proliferation can be measured by using a scintillation counter after elution to detect newly proliferated DNA cells that have incorporated the radiolabels. The downside of this method is that handling and using radioactive materials poses a serious risk. 

Analyzing the metabolic activity of a cell population

When the cell environment is metabolically active, the level of tetrazolium salts gradually decrease, resulting in the formation of a formazan dye that can alter the color of the growth medium. You can use a low or high profile spectrophotometer and a microplate reader to measure the metabolic activity of cells, which is an indicator of cell proliferation. 

Detecting ATP concentration in cells

The ATP content in cells can be detected using the bioluminescent luciferase and its substrate, luciferin. ATP is present only in living cells. Dead cells contain almost no ATP. The luciferase emits light only if ATP is present in cells and the intensity of the luminescence is directly proportional to the concentration of ATP in the cell. This technique is best suited for high-throughput cell proliferation assays and screening. 

Utilizing specific monoclonal antibodies

This technique is based on the presence of antigens in proliferating cells only. Antigens are absent in non-proliferating cells. This technique is widely used in cancer research for detecting the proliferation of tumor cells in vivo and in vitro. 

Additional resources

Regulation of cell proliferation by hypoxia-inducible factors

Cell Proliferation Assays

CytoTell™ Blue