Oxygen and the production of formazan from neotetrazolium chloride

Electrons, generated from dehydrogenase reactions, are transferred to oxygen in preference to neotetrazolium chloride. In model systems in solution the presence of a small amount of oxygen drastically reduces the rate of formazan production. The rate of reaction in tissue sections has been followed using scanning and integrating microdensitometry. As in solution, electrons are transferred preferentially to oxygen. However, oxygen seems unable to diffuse through the incubation medium and thus the supply of oxygen at the site of the enzyme activity becomes exhausted; the time taken to use up the oxygen will depend on the rate of the enzyme activity. It is only then that electrons are passed to the tetrazolium salt and formazan is precipitated.