Neutrophils isolated from human blood were labeled by various methods and exposed to a chemotactic gradient. The chemotactically functional cells that migrated into the gradient were isolated. The portion of radioactivity of the original cell suspension carried with the chemotactically responsive cells was related to the relative number of migrating cells as determined microscopically. Of the radionuclides used, P-32 diisopropylfluorophosphate (DFP), In-111 oxine, and Tc-99m sulfur colloid provided cell preparations with the highest relative portion of radioactivity confined to functionally intact (chemotactic) neutrophils. Results with Na2(51)CrO4 and with SnCl2-reduced 99mTcO4- were less than optimal. Neutrophils exposed to Ga-67 citrate apparently took up the label and retained chemotactic responsiveness. However, little or no radioactivity was detected in the neutrophils that migrated from the suspensions of Ga-67-labeled cells. The results indicate that the chemotaxis radioassay can yield unique information pertaining to the extent to which a radiotracer is specifically associated with viable neutrophils in a suspension of labeled cells.