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The surface monolayer technique was used to study the properties of biomembrane-like systems. The technique was used in combination with surface fluorescence measurements of fluorescent probes incorporated into a phospholipid matrix. Two types of fluorescent probe containing the chromophore pyrene bound to the hydrocarbon tail and the amino group of the phospholipid, respectively, were used. The fluorescent properties of the molecules were studied both in solution and in monolayers. On the basis of the results, fluorescence changes in the molecules upon compression of the monolayers studied arise from increased compactness in the phospholipid, which decreases permeation of atmospheric oxygen and hinders orientation changes in the chromophore at the interface.