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Quantitative determination of dehydrogenase activity in tissue sections and tissue homogenates.

The dehydrogenase activity in tissue sections was demonstrated by means of the ditetrazolium salt Nitro-Blue-Tetrazolium. It was found to be reduced to formazan compounds which, when precipitated in the tissues, indicate the enzyme activity. The same method can be employed for an estimatinon of the dehydrogenase activity in tissue homogenates. The formazan compounds are dissolved and the amount determined colorimetrically. The accuracy of the results depends on a complete dissolution of the produced formazan compounds. A method fulfilling these requirements is described. It was shown in a series of experiments that the produced amount of formazan indicates a corresponding enzyme activity (dependent on the added specific substrate). The reproducibility of the results is demonstrated and the influence of the pH values on the enzyme activity is discussed. The necessity of including control sections and blanks is illustrated. Comparative studies of semiquantitative (histochemical) and quantitative (colorimetric) measurements gave concordant results. Finally, it was shown that microscopy of tissue sections stained for dehydrogenase activity affords a possibility of estimating the intensities of the enzyme activities.