From the viewpoints of cellular morphology and enzymic histochemistry, we adopted the pseudopod method and NBT test for the measurement of spontaneous activation of neutrophils in vitro The time course curves of spontaneous activation of neutrophils from healthy donor were drawn. After comparing the pseudopod method with NBT test, we conclude that pseudopod method is more sensitive and reliable than NBT test and it can be used for real time observation; and that the curves of time course ascend from lower level to the highest level within 3 hours after the samples are collected, and then they descend gradually. The methodology of this study is available for use in future studies on the relationship between activation of neutrophils and many diseases.