Fluorescence microscopy and fluorescence redistribution after pattern photobleaching have been used to measure the distribution and motion of a number of fluorescent molecules bound to the plasma membranes of human leukocytes. The fluorescent molecules include fluorescein-labeled F(ab')2 and Fab' fragments of an anti-C3b receptor antibody, fluorescein-labeled IgG and Fab fragments of a monoclonal anti-HLA antibody, and the two lipid probes 3,3'-dioctadecylindocyanine and N-4-nitrobenzo-2-oxa-1,3-diazole L-alpha-dimyristoyl phosphatidylethanolamine. From these studies we have concluded that the C3b receptors on human polymorphonuclear leukocytes and monocytes are predominantly present in discrete clusters. No diffusive motion of these clusters could be detected. In contrast, HLA antigens hve a dispersed distribution and have diffusion coefficients between 10(-9) and 10(-10) cm2/sec in these peripheral blood leukocytes. Cell-membrane-bound monoclonal IgG anti-HLA undergoes a rapid patching in all these leukocytes except lymphocytes. In about 50% of the lymphocytes, fluoresceinated IgG diffuses slowly, if at all, whereas in the other 50%, fluoresceinated IgG diffuses with a diffusion coefficient of 6.9 X 10(-10) cm2/sec. The two above-mentioned fluorescent lipid probes have the usual rapid lipid diffusion coefficient (approximately 10(-8) cm2/sec) in leukocytes.