Track-A-Worm (version 2.0)

Version 2 now comprises three components: WormTracker, SleepTracker, and Action Potential Analyzer. The WormTracker represents an enhanced iteration of the original Trak-A-Worm, featuring substantial improvements and novel functionalities. Below is a list of the major additions and enhancements:

  1. Option to differentiate between ventral and dorsal sides.
  2. Implementation of image recording in greatly reduced file sizes (due to a change from BMP to JPG format), thereby conserving storage space and expediting subsequent analysis.
  3. Implementation of body curvature quantification.
  4. Capability to track and analyze worms on nematode culture plates with a layer of OP50 or other bacteria.
  5. Automatic spline fitting for virtually all worm shapes.
  6. Option to integrate external devices (g.,a light source for optogenetic stimulation) through TTL signals.
  7. Plotting of forward and backward locomotion profiles.
  8. Resolution of all bugs identified in the original version.

    Instructions for system setup

    Software installation (Windows OS)

    1. Download the free Matlab Compiler (R2021b, 64 bit for Windows). This step may be omitted if you already have Matlab installed on your computer.
    2. Download the Track-A-Worm v2 (a compressed file).
    3. Decompress the installer and install it.

    Hardware setup


    Video tutorials

    1. Camera Calibration
    2. Record
    3. Playback
    4. Fit Spline
    5. Analyze
    6. Batch Spline
    7. Batch Analyze
    8. Curvature Analysis
    9. Sleep Recorder
    10. Sleep Analyzer
    11. Action Potential Analyzer

    User Manual for v2.0



    This MATLAB-based app allows you to correct the drop in fluorescence signal over time due to photobleaching of a genetically encoded calcium sensor such as GCaMP6. The software uses the cubic interpolation algorithm to create a curve along the baseline chosen by the user.  Subsequently, the raw data are divided by the interpolated points of the baseline to generate the normalized data. The software may be used by following these steps: 1) Assign regions of interest (ROIs) in your calcium imaging data using ImageJ (National Institute of Health); 2) plot fluorescence intensities (F) in each ROI as absolute intensities over time; 3) save only the fluorescence intensities as a text file; 4) run the app by clicking the "baseline.m" file; 5) load the text file into the app to produce a trace of the raw data; 6) click baseline points along the plot while holding down the "Alt" key, followed by clicking "Done"; 7) Click "Yes" if the spline (red color) generated by the app represents the baseline well; 8) save the data (F/F0). Click baseline to download the app.