Knowledge Base

A Conditioning Test in EthoVision XT

Create an experiment. For details, see Chapter 1 of this manual, or in EthoVision XT press F1 and see Setup an Experiment in the EthoVision XT Help. In the following sections we assume that you created an experiment using the PhenoTyper arena template.

Experiment Settings

1. Make sure that the PhenoTyper cages are connected to the EthoVision XT PC. See the PhenoTyper - EthoVision XT 18 - Reference Manual for how to connect PhenoTyper to EthoVision XT. Connect a third-party hardware device, which can be controlled by TTL, to one of the TTL control ports on the USB-IO Box.
2. Open the experiment and choose Setup > Experiment Settings.
3. Check that the correct cameras are selected under Video source. Select whether the resolution of the first camera and that of the mixed image. Note that depending on the number of cameras and the frame rate, you may need to reduce the resolution of the mixed image. For more information about installing and connecting multiple cameras, see Camera installation in the EthoVision XT Help and the PhenoTyper - EthoVision XT 18 - Reference Manual.
4. If you want to use the Deep learning technique to track the subject's nose, under Body point detection technique choose Deep learning. Note that in order to use this technique there are additional requirements. Among other things, you need a powerful graphics card. See Deep learning: Requirements and Limitations in the EthoVision XT Help.
5. Under Analysis options, select Activity analysis if you want to measure freezing behavior.
6. Next to Hardware, select Use of Trial Control Hardware and click Settings.
7. Select Noldus USB-IO box.
8. In this next step, you define the ports of the USB-IO box connected to PhenoTypers. Select SDI port 13, then Top Unit Interface. Select Top Unit (Standard) for all the rows below (depending on how many PhenoTypers are connected to a Top Unit Interface).

   For PhenoTyper 2, and for more details and examples, see the PhenoTyper - EthoVision XT 18 - Reference Manual.

   If you use third-party devices, also select the TTL ports that are connected to those devices.

9. Click OK.

For More Information

• About port mapping: the EthoVision XT 18 - Trial and Hardware Control - Reference Manual.
• About PhenoTyper connections and devices:
  • PhenoTyper - EthoVision XT 18 - Reference Manual
  • PhenoTyper - EthoVision XT 18 - Service Manual for specific devices like the Pellet dispenser and the Lickometer.
• About general experiment settings: Set Up an Experiment in the EthoVision XT Help.

Arena Settings

Choose Setup > Arena Settings > open Arena Settings 1.

Click the Arena - Hardware Mapping button at the bottom of the Arena Settings pane.

If you only have one arena, the hardware will be assigned to this arena automatically. If that is the case, proceed with Trial Control Settings.

In the Arena - Hardware Mapping window, click Add Device for each device you want to add:

• For PhenoTyper: Under Device Type, select Top Unit (Standard). Enter Device Name PhenoTyper and the corresponding Top Unit (Standard) # for each Arena.
• For a third-party hardware device: Under Device Type, select Custom Hardware. Enter a Device name and select a device from the list for each Arena.

Click 4. Validate Setup to validate the Arena Settings.

Trial Control Settings

With the Trial and Hardware Control add-on module in EthoVision XT, you can control PhenoTyper and third-party devices in a variety of ways. For example:

• Start recording when the animal is detected on top of the shelter.
• Stop recording when the animal has performed a specific task 50 times.
• When the animal sits on the shelter, drop a food pellet. Repeat this task 25 times.

The template experiment contains two trial control profiles:

• Default – The default Trial Control Start-Stop rule.
• Max. Trial duration 24 hrs – In this profile, a trial stops after 24 hours.

Maximum trial duration (tested):

• Without video recording: 72 hours.
• With simultaneous video recording: 3 hours.

If you need to record video of the same subject for more than three hours, we recommend that you make a series of trials.

Below you find a few examples of Trial Control Settings. Modify the Trial Control Settings to suit your own application.

Procedure Overview

1. The trial starts with a 30-second delay (see Start Trial Rule and Condition Start Delay below).
2. Next, as soon as the mouse enters the LightSpot zone, with both the nose- and center-point, the spotlight of the PhenoTyper is turned on. When both the nose- and center-point are not in the LightSpot zone, the spotlight is turned off. This sequence is repeated three times (see Sub-rule Activation LightSpot below).
3. After the Sub-rule is completed, the trial is stopped after a 5-second delay.

Choose Setup > Trial Control Settings > New and give this a new name.

Start Trial Rule

1. To start recording as soon as the Start trial button is clicked:
   • Click the (default) Condition In zone box and press Delete.
   • Keep this Condition if you want to start recording some time after starting the trial.

Condition 'Start Delay'

1. To insert a 30-second delay after recording has started:
   1. In the Components pane, under Conditions, double-click Time Condition or click the button next to it.
   2. In the Time condition window, type Delay of 30 seconds in the Condition name field. In the Condition is met group, select After a time of 30 secs. Optionally, enter a comment and click OK.
   3. Insert the Condition box into the main sequence.

For more information about Conditions, see the EthoVision XT 18 - Trial and Hardware Control - Reference Manual.

Sub-Rule 'Activation LightSpot'

1. Because the sequence of turning on/off the spotlight depending on whether the animal is inside/outside the Spot Light Zone is repeated three times, you need to create a Sub-rule for this:
   1. In the Components pane, under Structures, double-click Sub-rule or click the button next to it.

Source: EthoVision XT 18 - Application Manual, Noldus Information Technology