In the computer, open MATLAB. Add the folder containing the MouseWalker script to the working directory and run MouseWalker. M on the main command line.
Load the video folder as the input directory. Go to the Settings window where all the calibration and threshold parameters are located. Test the effect of changing some parameters by clicking the Preview button.
After adjusting the threshold parameters, check that the video is ready for automated tracking. Go to the first frame and click auto to start tracking. After the tracking is completed, if required, perform a manual correction by selecting the appropriate footprint.
Save the changes by pressing the Save button. Then, click evaluate to generate the output files from the tracked video. Check that all the graphical output data plots are saved in the Results folder.
Next, check that all quantitative measurements generated by the MouseWalker software are saved on an Excel spreadsheet and summarized on 1.Information_Sheet. Use the MouseMultievaluate. M script to congregate the measurements from all the runs into a new file for analysis.
To perform a principal component analysis, PCA, open pcaplotgenerator. py in Spyder and run the code by clicking on the Play button. Select the Excel file to analyze and the sheet name in the automatic window.
If the sheet name is not altered, write Sheet 1. In the digital ink assay, a lack of support for the hind paws was detected. With a decrease in the footprint area for both the left and right hind paws.
The overall stance traces displayed several unique features. After spinal cord injury, the hind paws had shorter stance traces and more random pore positioning at both touchdown and liftoff from 15 days post-injury. Principal component analysis of all the kinematic motor parameters showed a 40%variance in the data in the first component, which segregated the group of animals that had a spinal cord injury from the rest at all time points.
Using other scripts such as the heat map script, it was found that the spinal cord injury animals showed alterations in the gait strategies at all time points. The spinal cord injury mice also displayed a lower stance straightness index in both the forelimbs and hindlimbs.