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To evaluate the psychometric properties of 4 measures of acute pain in youth with sickle cell disease (SCD) during a medical procedure.Heart rate, child self-report, parent proxy-report, and observable pain behaviors were examined in 48 youth with SCD ages 2 to 17 years. Criterion validity for acute pain was assessed by responsiveness to a standardized painful stimulus (venipuncture) in a prospective pre-post design. Convergent validity was evaluated through the correlation across measures in reactivity to the stimulus.Child self-reported pain, parent proxy-report, and behavioral distress scores increased in response to venipuncture (concurrent and convergent validity). In contrast, heart rate did not reliably change in response to venipuncture. Extent of change in response to venipuncture showed moderate intercorrelation across child and parent pain ratings, and behavioral distress. Preprocedure pain ratings correlated with pain experienced during the procedure. An item analysis of observable pain behaviors suggested differences in the presentation of pain in SCD compared with previous pediatric research.Criterion and convergent validity were demonstrated for child-report, parent-report, and observable pain behaviors. These measures seem to tap into distinct, yet overlapping aspects of the pain experience. Assessment of acute procedural pain responses in SCD requires evaluation of preprocedural pain due to the frequent presence of low-level, baseline pain.