We developed a method for determining whether channels in a multichannel patch or bilayer have the same or statistically significantly different open probabilities. We use a maximum likelihood method to fit the distribution of (unbinned) current amplitudes and to provide estimates of individual channel open probabilities, single channel currents, and standard deviations of the channel currents. These parameters are used to compare models with increasing constraints on the open probabilities including the model where all channels have different open probabilities and the model where all channels have the same open probability. A chi 2 statistic is used to identify models that are statistically less likely to predict the data. The ability of multichannel data to determine individual open probabilities is limited by two factors: the signal to noise ratio of the record and the fact that changes in amplitude distributions caused by a 0.2 difference in open probabilities are comparable in magnitude to the variations caused by random channel gating. These limitations notwithstanding, we demonstrate the utility of our approach by using it to analyze the open probabilities of 3 large conductance Ca2(+)-activated K+ channels in an artificial lipid bilayer revealing the response of one of those channels to GTP gamma S.