Yeungnam Univ J Med.  2018 Jun;35(1):63-69. 10.12701/yujm.2018.35.1.63.

Reliability and validity of free software for the analysis of locomotor activity in mice

  • 1Department of Pediatrics, College of Medicine, Kosin University, Busan, Korea.
  • 2Department of Psychiatry, Pusan National University Hospital, Busan, Korea.
  • 3Department of Psychiatry, Pusan National University School of Medicine, Busan, Korea.


Kinovea software that tracking semi-automatically the motion in video screen has been used to study motion-related tasks in several studies. However, the validation of this software in open field test to assess locomotor activity have not been studied yet. Therefore, this study aimed to examine the reliability and validity of this software in analyzing locomotor activities.
Thirty male Institute Cancer Research mice were subjected in this study. The results examined by this software and the classical method were compared. Test-retest reliability and inter-rater reliability were analyzed with Pearson's correlation coefficient and intraclass correlation coefficient (ICC). The validity of this software was analyzed with Pearson's correlation coefficient.
This software showed good test-retest reliability (ICC=0.997, 95% confidence interval [CI]=0.975-0.994, p < 0.001). This software also showed good inter-rater reliability (ICC=0.987, 95% CI=0.973-0.994, p < 0.001). Furthermore, in three analyses for the validity of this software, there were significant correlations between two methods (Pearson's correlation coefficient=0.928-0.972, p < 0.001). In addition, this software showed good reliability and validity in the analysis locomotor activity according to time interval.
This study showed that this software in analyzing drug-induced locomotor activity has good reliability and validity. This software can be effectively used in animal study using the analysis of locomotor activity.


Locomotor activity; Video-tracking software; Reliability; Validity; Open field test

MeSH Terms

Motor Activity*
Reproducibility of Results*


  • Fig. 1. Classical method measures locomotor activity in the open field test by counting the number of times that the forelegs of the mouse pass crossed lines (A), and Kinovea method measures locomotor activity by tracking movements of the mouse (B).

  • Fig. 2. This graph showed good test-retest reliability of Kinovea tracking software program in two times analyses at a month-interval (ICC=0.988 [95% CI=0.975-0.994], p<0.001). ICC, intraclass correlation coefficients; CI, confidence interval.

  • Fig. 3. This graph showed good inter-rater reliability of Kinovea tracking software program through the same video clip by two persons in two times analyses (ICC=0.987 [95% CI=0.973- 0.994], p<0.001). ICC, intraclass correlation coefficients; CI, confidence interval.

  • Fig. 4. This graph showed good validity of Kinovea tracking software program compared to classical method in order to assess locomotor activity in open field test in two times analyses (first analysis: Pearson's correlation coefficient=0.928, p<0.001; second analysis: Pearson's correlation coefficient=0.942, p<0.001; third analysis: Pearson's correlation coefficient=0.972, p<0.001).


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