The purpose of the present study was to evaluate moderate-to-vigorous physical activity using triaxial accelerometry in Japanese preschool children. The relationship between daily step counts as a convenient measure of physical activity and minutes of engagement in moderate-to-vigorous physical activity was also examined. Physical activity was assessed using a triaxial accelerometer (ActivTracer, GMS) and daily steps using a uniaxial accelerometer for 6 consecutive days, including weekdays and weekend days, in 157 four- to six-year-old Japanese children attending kindergarten or nursery school. Using triaxial and uniaxial accelerometers, nonlocomotive activities and step counts for young children can be evaluated, respectively. Average daily moderate-to-vigorous physical activity (physical activity ratio ≥3) and step counts were 102 (±32) min/day and 13,037 (±2,846) steps/day, respectively. A strong and significant correlation was observed between minutes of moderate-to-vigorous physical activity and step counts (r=0.832, p<0.001). The daily step counts corresponding to 60 min, 100 min, and 120 min of moderate-to-vigorous physical activity were 9,934, 12,893, and 14,373 steps/day, respectively. The correlation coefficient between minutes of higher intensity activities (physical activity ratio ≥4) and step counts was slightly lower (r=0.604, p<0.001). The daily step count corresponding to 30 min of the higher intensity activities was 14,768 steps/day. These results suggest that approximately 13,000 steps/day are required for preschool children to engage in more than 100 min of moderate-to-vigorous physical activity.