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        MTT Assays

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        The MTT calorimetric assay determines the ability of viable cells to convert a soluble tetrazolium salt [3–(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) into an insoluble formazan precipitate. Tetrazolium salts accept electrons from oxidized substrates or appropriate enzymes, such as NADH and NADPH. In particular, MTT is reduced at the ubiquinone and cytochrome b and c sites of the mitochondrial electron transport system and is the result of succinate dehydrogenase activity. This reaction converts the yellow salts to blue-colored formazan crystals that can be dissolved in an organic solvent whose concentration can be spectrophotometrically determined (Fig. 1 ). Owing to the many advantages of the assay, it is today considered a significant advance over traditional techniques. In fact, it is rapid, versatile, quantitative, and highly reproducible with a low intratest variation between datapoints (�15% SD); it is useful in a large-scale, antitumor drug-screening program (1 3 ). Moreover, the test can also be used for floating cells, such as leukemias and small cell lung carcinoma, and always allows sufficient time for cell replication, drug-induced cell death, and loss of enzymatic activity, which generates the formazan product from the MTT substrate (4 ).
        Fig. 1.  Absorption spectra of MTT formazan reagent (25 μg/mL) in DMSO (A ) and 0.04N HC1/isopropanol (B ).

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