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In 1941, Kuhn and Jerchel1 reported, citing the pioneering observations of several authors including those of themselves, that tetrazolium salt, a colorless compound, is reduced to the red formazan by the enzymatic activity of living cells. Here is a very rare example in organic chemistry that the reduced state of a dye is colored and the oxidized one is colorless. In addition, this red substance has convenient properties that it is stable against oxygen, water insoluble, but soluble in organic solvents such as aceton, ether and so on. Later studies on this phenomenon presented an additional support for the applicability of tetrazolium salts for many kinds of experimental purposes in biology and medicine. Black and his co-workers2, 3, 4 have engaged in their cancer research employing 2, 3, 5 triphenyltetrazolium chloride (TTC). Mac Kenzie and Fuller5, and Schuermann6 also used this substance as an indicator for malignant tumors. Some others7,8 attached their interest to the study of cell metabolism by the tetrazolium reduction phenomenon. These investigations are based in principle upon the dehydrogenase activity of cells measured by the amount of formazan produced by them. On the other hand, the experiments by Mudd et al9, and Burns and Militzer10 suggested that the localization of TTC reaction in bacteria may be the mitochondoria. Now, the recent paper of Mac Vandiviere11 snowed that the total viability of a suspension of tubercle bacilli could be estimated by photometric measurement of the optical density of aceton solution of the reduced TTC produced by living tubercle bacilli. This report stimulated us considerably and led to the attempt to confirm and extend his observation. The present paper deals chiefly with the fundamental aspects of the techniques of the TTC reduction test against tubercle bacilli, presenting the data obtained in the following experiments which were made for the purpose of detecting the most suitable condition for the test.

TETRAZOLIUM REDUCTION TEST AS A MEASURE TO EXAMINE THE TOTAL VIABILITY OF SUSPENSIONS OF TUBERCLE BACILLI I. STUDIES ON THE TECHNIOUES OF THE TEST