Derick Mussen Preventive Healthcare

Nitroblue Tetrazolium Slide Test

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This protocol is a simple method for assessing the ability of activated neutrophils to generate reactive products of oxygen. It provides a rapid, albeit qualitative, determination of NADPH oxidase activity. Neutrophils in a drop of freshly obtained blood are allowed to adhere to a stimulus-coated coverslip and then incubated with nitroblue tetrazolium (NBT). Products of the respiratory burst, primarily superoxide, reduce the NBT to formazan, which forms a blue-black precipitate in the positive cells. Individual cells are scored microscopically for positive NBT reactions.

Materials

Stimulus: 500 ^g/ml endotoxin or 10 ^g/ml phorbol myristate acetate (PMA) Nonanticoagulated blood (e.g., from a fingerstick)