Jan 10, 2025
Catwalk Gait Analysis Protocol
- 1University of Alabama at Birmingham
Protocol Citation: Ashley Adamson, Allie Smith, Briana R De Miranda 2025. Catwalk Gait Analysis Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.5jyl8dp7rg2w/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: December 07, 2024
Last Modified: January 10, 2025
Protocol Integer ID: 114528
Funders Acknowledgements:
Briana De Miranda
Grant ID: R01ES034846
Abstract
Motor and gait impairments are important outcome measures for many disease phenotypes. The following is a protocol for assessing motor and gait behavior using the Catwalk XT gait analysis platform at the University of Alabama at Birmingham in the Neuroscience Behavioral Assessment Core.
Guidelines
Use of this protocol needs approval by user's Institutional Animal Care and Use Committee (IACUC) or equivalent ethics committee.
Materials
- Catwalk XT version 10.6 platform and software
- Chlorhexidine
- Paper towels
- Test mouse
- Experimental mice
Before start
You will need a test mouse whose behavior is not included in the experiment to calibrate detection settings
Set Up
Set Up
Bring animals into testing room for habituation (at least 30 minutes prior to start of experiment)
Turn on power switch
Clean walkway with chlorhexidine
Turn down lights
Darker lights increase the sensitivity of the platform
Make note of light settings so that you can remain consistent across runs
Open the Catwalk XT Version 10.6 program
Set up the walkway
Camera should be 35 cm away for mice or 70 cm for rats
Define the walkway – drag the box to take up the whole width and length of the catwalk that is within the camera view
Calibrate walkway – place a calibration paper down on the walkway and drag the square in the program the same size
- Tell the program how big the square is based on the known size of your calibration paper
Create new experiment
Name it
Indicate where the file needs to be saved
Experiment settings
Choose mouse or rat based on what type of animal is being tested
Input the groups that you have (i.e. control, experimental, sham, etc)
Criteria:
- Minimum run time = 0.5 seconds
- Maximum run time = 5.0 seconds
- Runs = 3
- Maximum variation = 60%
Trial List
Add the number of trials equivalent to the number of animals being tested
Optional: add the trials to defined groups to separate out experimental groups
Detection settings
Located inside setup tab
Gain: 220-225
Place a test mouse inside the corridor
Start auto detection
Then adjust the intensity threshold to clear out noise
- Look at prints to ensure that only the feet are registering (and no other body parts)
Acquisition settings
Located inside setup tab
Auto abort at 10 sec
Discard auto aborted runs
Discard runs noncompliant with run criteria
Discard incomplete prints at the start and end of a run
Optional: toggle on/off sound feedback as desired
Running the Experiment
Running the Experiment
Aquisition
Select the desired trial
Snap background image
Click start aquisition
Place mouse in corridor
Run for a total of 3 compliant runs for each animal
Exlcude mice after 30 failed runs
Repeat for all mice
Analysis
Analysis
Classify Runs
Select trial and run
Auto classify
Use the “UN ->” and “<- UN” to navigate through unclassified prints
Click and drag over UN to identify the unclassified prints
If UN is not a print, select the print and classify as junk
Repeat for all compliant runs
Visualization
Make sure all date and all parameters are selected on the left
Scroll through the run to visualize the prints
Run stats
Illustrates both the print view and the temporal view of each run
Lists the statistic broken down by parameter for each run
View Group Statistics
Shows the data broken down by group (if groups were used)
Export
File -> export -> trial statistics
Chose file type
Automatically saves in the export folder
Acknowledgements
We would like to thank the UAB Neuroscience Behavioral Assessment Core for use of their facilities.