The purpose of this study was to compare vastus medialis obliquus:vastus lateralis muscle (VMO:VL) integrated electromyographic (IEMG) ratios of healthy subjects and patients with unilateral patellofemoral pain (PFP) under isotonic and isometric quadriceps femoris muscle contraction conditions. Subjects ranging in age from 18 to 35 years (mean = 28.06, SD = 5.97) were assigned to one of three groups on the basis of type of knee condition. In group 1, which consisted of seven healthy control subjects with no history of knee pathology, both knees were tested. In group 2, which consisted of nine patients with unilateral PFP, only the painful knee was tested. In group 3, which consisted of the same nine patients who comprised group 2, only the nonpainful knee was tested. Nonnormalized and normalized VMO:VL IEMG ratios were computed for ascending stairs, descending stairs, submaximal isometric contraction, and maximal isometric contraction (nonnormalized only). A two-way analysis of variance for repeated measures indicated VMO:VL ratios for isotonic stair-climbing activities were significantly greater than VMO:VL ratios for isometric contractions. Nonnormalized VMO:VL ratios in group 1 were significantly greater than nonnormalized VMO:VL ratios in the other two groups. Patients with PFP may have abnormal VMO:VL activation patterns, and isotonic quadriceps femoris muscle exercise may elicit more favorable muscle activation patterns than isometric exercise for patients with PFP.