This tutorial demonstrates how to conduct a Y-maze protocol using adult zebrafish in the Zantiks AD unit.
AD set up
A Y-maze insert will be used to create a testing environment for two zebrafish.
Subjects do not need extensive single-housing beforehand. Approximately 30 minutes before testing, subjects can be moved to individual tanks. With two arenas per tank, two fish can be tested concurrently in each unit. The experiment is conducted in the dark to reduce stress to the fish.
Before beginning the service (the experimental script), you can check that the fish are freely moving in each arena by opening the live video page from the unit's console.
Setting up the Zantiks AD for the Y-maze study with adult zebrafish
The following script is an example of one used in a typical Y-maze study.
Whilst familiarising yourself with scripting, it is advisable to use the script below in tandem with the Zanscript syntax manual.
ACTION MAIN DEFINE TRIAL_TIME 3600 SET(TARGET_SIZE,5) SET(DETECTOR_THRESHOLD,3) SET(AUTOREF_MODE,MOVEMENT) SET(AUTOREF_TIMEOUT,30) LOAD(ARENAS,"arenas_y_maze_2") LOAD(ZONES,"zones_y_maze_2") LOAD(ZONECHANGES,"on") AUTOREFERENCE() WAIT(TRIAL_TIME) COMPLETE
- The trial time is defined as one hour (3600 seconds).
- Four SET commands tell the unit the tracking settings for the adult zebrafish.
- The next two LOAD commands tell the unit which assets to use for the arenas and zones.
- The LOAD(ZONECHANGES,"on") command tells the unit to record every zone change into the data log file.
- The unit then waits for the time defined above as TRIAL_TIME. The unit then waits the time defined above as TRIAL_TIME. The zone changes will therefore be recorded for this time period.
Test data results
The results from the Y-maze experiments will be saved into the Data directory as a .csv file. From these results it is possible to obtain data detailing the sequential arm visits, which can then be analysed as a tetragram, triad, or similar.
One way to analyse the data can be found on github. Here you will find instructions and an R script which is designed to batch analyse raw data files created by the Zantiks unit into tetragrams.
See the featured protocol page, Y-maze (zebrafish, Drosophila, mice), for more details on the assay and script downloads to run the assay.
You will need to upload the asset into the Asset directory on your Zantiks Control Console and ensure the correct asset name is in the LOAD(DETECTORS,"name_of_asset") command in the script.