Subclasses of opiate receptor binding sites in human brain membranes were investigated by means of competitive binding techniques. The experimental data were analyzed by use of a computerized non-linear regression curve fitting program. mu-, delta-and chi-types of opiate binding were found in 5 different regions of the brain. A more extensive analysis of the regional distribution of subclasses of opiate binding sites was performed using a simple sequential inhibition technique. This method was shown to yield results which are comparable to those obtained by computer analysis of multiple tracer displacement curves. Chi-and mu-sites represented the major component of binding in most brain areas whereas delta-sites were fewer in number. The 3 types of binding showed different distribution patterns, suggesting that they are independent from each other. The distribution pattern observed in human brain resembled the one observed in rat brain, although chi-sites appear to represent a more important, and delta-sites appear to represent a less important, fraction of binding in human as compared to rat brain.