Apr 28, 2026

Open Field Exploration: Locomotor Activity and Anxiety Assessment in Mice

DOI
https://dx.doi.org/10.17504/protocols.io.rm7vz4d2rlx1/v1
  • 1KU Leuven;
  • 2Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD 20815
Sarah van Veen
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DOI: https://dx.doi.org/10.17504/protocols.io.rm7vz4d2rlx1/v1
Protocol Citation: Sarah van Veen, Zsuzsanna Callaerts-Végh 2026. Open Field Exploration: Locomotor Activity and Anxiety Assessment in Mice. protocols.io https://dx.doi.org/10.17504/protocols.io.rm7vz4d2rlx1/v1
License: This is an open access  protocol  distributed under the terms of the  Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: April 21, 2026
Last Modified: April 28, 2026
Protocol  Integer ID: 315426
Keywords: ASAPCRN, anxiety assessment in mice, assessment of locomotor activity, indices of locomotor activity, behavior in adult mice, locomotor activity, anxiety assessment, open field exploration, anxiety, mice, adult mice, mice this protocol, field arena
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: ASAP-000458
Fonds Wetenschappelijk Onderzoek (FWO)
Grant ID: G011424N
Abstract
This protocol describes assessment of locomotor activity and anxiety-related behavior in adult mice using
an open-field arena. Total path length and corner crossings serve as indices of locomotor activity; entries
into the centre serve as a measure of conflict resolution or anxiolysis.
Guidelines
•       Testing order should be counterbalanced or randomized across genotypes.
•       All testing should be performed at the same time of day across animals.
•       Experimenter should leave the room or remain still/silent during the trial.
Materials
AB
SubjectsAdult WT and KO littermates (~2 months old)
Arena50 × 50 cm2 square open-field arena
TrackingANY-maze video tracking equipment and software (Stoelting Europe, Ireland)
Cleaning70% ethanol solution + absorbent wipes
Dark habituationDark room or light-proof box for pre-trial habituation
Before start
•       Allow mice to habituate to the procedure room for ≥30 min before testing.
•       Prepare 70% ethanol and wipes for inter-animal cleaning.
•       Test the recording and tracking system before the first animal.
Place each mouse in a dark holding space (or covered holding cage) adjacent to the test room for 30 min immediately before testing.
Transfer the mouse gently to a corner of the open-field arena. Start ANY-maze recording immediately. Allow free exploration for 10 min.
Record behavior for 10 min
ANY-maze automatically tracks position and computes: (a) total path length (cm) and corner
crossings — locomotor activity indices, which are highly correlated and mainly indicate thigmotactic
walking near the walls of the arena; (b) entries into the centre — recorded as a measure of conflict
resolution or anxiolysis.
After 10 min, return the mouse to its home cage. Thoroughly clean the arena floor and walls with 70% ethanol. Allow ethanol to fully evaporate before placing the next mouse.
Export total path length, corner crossings, and centre entries for each animal from ANY-maze. Apply
appropriate statistical tests comparing WT vs KO.