Apr 28, 2026

DigiGait: Quantitative Gait Analysis in Mice

DOI
https://dx.doi.org/10.17504/protocols.io.n92ld4xn9l5b/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.n92ld4xn9l5b/v1
Protocol Citation: Sarah van Veen, Zsuzsanna Callaerts-Végh 2026. DigiGait: Quantitative Gait Analysis in Mice. protocols.io https://dx.doi.org/10.17504/protocols.io.n92ld4xn9l5b/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: 315435
Keywords: ASAPCRN, quantitative gait analysis in mice, gait analysis in adult mice, automated gait analysis, quantitative gait analysis, multiple spatiotemporal gait parameters during treadmill locomotion, multiple spatiotemporal gait parameter, treadmill locomotion, mice this protocol, use of the digigait system, adult mice, digigait system, mice, digigait, plane imaging
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: ASAP-000458
Fonds Wetenschappelijk Onderzoek (FWO)
Grant ID: G011424N
Abstract
This protocol describes the use of the DigiGait system (Mouse Specifics Inc.) for high-speed ventral-plane imaging and automated gait analysis in adult mice. The system quantifies multiple spatiotemporal gait parameters during treadmill locomotion.
Guidelines
•       Use the same belt speed for all animals in a cohort.
•       Ensure the imaging surface is clean — debris causes tracking artifacts.
•       Perform analysis blinded to genotype where possible.
Materials
AB
SubjectsAdult WT and KO littermates (~2 months old)
ApparatusDigiGait imaging system (Mouse Specifics Inc., Framingham, Massachusetts, USA)
CameraHigh-speed ventral-plane camera (integrated)
Treadmill belt speedstarting speed 10 cm/s, increasing to 15 cm/s once animal is walking steadily
SoftwareDigiGait Analyzer software
Session duration~5–10 s of steady-state locomotion per animal
Before start
•       Start DigiGait system and DigiGait Analyzer software ≥15 min before use to allow warm-up.
•       Set treadmill belt speed appropriate for the study.
•       Clean treadmill belt and transparent walking surface with 70% ethanol before each animal.
•       Allow mice to acclimate to procedure room for ≥30 min before testing.
Animal acclimation
Gently place mouse on the stationary belt with the enclosure door closed. Allow 1–2 min for the animal to explore and calm before initiating belt movement.
Initiate belt movement
Start the treadmill at 10 cm/s, then increasing to 15 cm/s once the animal is walking. Allow the mouse to reach steady-state locomotion (consistent running without stumbling or stopping). This typically takes 10–30 s.
Record gait
Once steady-state locomotion is achieved, begin video capture for at least 5 full stride cycles (aim for 5–10 s of continuous recording). The ventral camera records paw placement for all four limbs simultaneously.
Return mouse to home cage
Stop belt, open enclosure, and return mouse to its home cage. Clean belt surface with 70% ethanol before next animal.
Automated analysis
Load video into DigiGait Analyzer. The software automatically segments individual paw areas across frames and computes gait parameters including: stride length, stride frequency, stance width (front/hind), paw area, swing time, stance time, and symmetry indices.
Data export and statistics
Export gait parameters per animal. Compare WT vs KO groups using appropriate tests.