The purpose of the current study was to investigate how combinations of different epoch durations and cut-points affect the estimations of sedentary time and physical activity in adolescents. Accelerometer data from 101 adolescents were derived and 30 combinations were used to estimate sedentary time, light, moderate, vigorous, and combined moderate-to-vigorous physical activity. Data were analyzed with repeated measurement analyses of variance. Large differences of sedentary time and times of different physical activity intensities were observed between 1 s and longer epoch durations using virtually all cut-points. Generally, sedentary time, moderate physical activity, vigorous physical activity, and combined moderate-to-vigorous physical activity progressively decreased, whereas light physical activity increased with longer epoch durations. The extreme differences between cut-points were large and increased with longer epoch durations for sedentary time and for all physical activity intensities except for vigorous physical activity per epoch duration. Caution is required when cross-comparing studies using different epoch durations and cut-points. To accurately register adolescents' spontaneous intermittent physical activity behavior, short epoch durations are recommended.