Knowledge Base

Experiment in EthoVision XT

Prerequisites

Make sure that all devices are connected and powered up.

Create an EthoVision XT Experiment

1. Start EthoVision XT and choose File > New From Template, then click Apply a pre-defined template.
2. On the How is the arena configured page, select:
   • If you work with one or two cubicles, Fear Conditioning System cubicle. Next to Number of arenas, select the appropriate number.
   • If you work with four cubicles, Fear Conditioning System cubicles. Next to Number of arenas, select the appropriate number.

   Note: This template does not include pre-defined zones as they are not used often in Fear Conditioning.

3. When asked How many subjects per arena will you track, select subject.
4. Complete the guided procedure and give the experiment a name. Click OK.

Experiment Settings

Cameras

1. Choose Setup > Experiment Settings.
2. Under Video Source, next to Number of sources, select how many cameras (one per cubicle) you are working with. In the list under Source, select the name of each camera.

   The IP address should indicate which camera belongs to which cubicle, provided that you have assigned IP addresses accordingly.

3. Next to Merged camera view, select 800 x 600 (cropped view of the cage) or 1280 x 1024 (full view of the cage).

Number of Arenas and Tracked Features

Important: Do this before selecting the Ugo Basile FCS in the Trial Control Hardware Settings.

1. In the Experiment Settings, under Arenas, select the Number of Arenas you use, that is, the number of cubicles (1 to 4).
2. Under Tracked Features, in most cases Center-point detection will suffice for fear conditioning experiments.

Activity Analysis for Freezing Detection

1. In the Experiment Settings, under Analysis Options, select Activity analysis if you want to analyze freezing behavior.

   Tip: Click the Video Tutorial button next to that option for an overview of Activity analysis.

Specify the Ugo Basile Fear Conditioning System

Important: Ensure you have already selected the number of arenas. Make sure that Number of Arenas selected corresponds to the number of FCS cages used in that experiment. Only after that, follow the instructions below.

1. In the Experiment Settings, under Trial Control Hardware, select Use of Trial Control Hardware, and click Settings.
2. In the Hardware Interface window, select Ugo Basile F.C. Controller.
3. In the Fear Conditioning System COM Port Settings window, click the Add button.
   • Select a COM port and click OK. Repeat this step to add the COM ports connected to all the FCS cubicles.
   • For a setup with the Ugo Basile USB Hub and four cubicles, add COM11, then COM12, COM13, and COM14.
   • For a setup with one cubicle connected directly to the PC, select the COM port associated with that cubicle. If you have created the experiment from a template, there may be one or more COM ports already selected in the Fear Conditioning System COM Port Settings window. If they do not match those associated with the cubicles of your setup, delete them and add the correct ones.
4. When finished, click OK and proceed with arena settings.

Important Notes:

• Take note of which cubicle is connected to which COM port.
• Make sure that only the necessary COM ports are added. If you accidentally added a wrong COM port, select it and click Delete.
• When adding more COM ports in the Fear Conditioning System COM Port Settings window, start with the port for cubicle 1. Disconnect cubicle 1 and see which COM port disappears from the Windows Device Manager. Reconnect the cubicle and add that COM port. Repeat those steps for cubicles 2, 3, and 4.

COM Port Communication

To determine which COM port is associated with a specific cubicle, do the following:

1. In Windows, open the Device Manager. Choose View > Devices by type.
2. Open the item named Ports (COM and LPT).

   If the cubicles are connected physically to the PC through the Ugo Basile USB Hub and powered up, you should see one to four items labeled USB Serial Port (COMx): COM11 to COM14.

   Tip: If you do not see the Ports item, from the View menu of the Device Manager select Devices by type.

3. Disconnect the USB cable from one of the FCS cubicles.
4. Look at the Ports (COM and LPT) list. The item that has disappeared is the COM port currently associated with that cubicle.

   Tip: Number the FCS cages and cubicles, and take note of which COM port is linked with each cubicle. This makes troubleshooting easier.

   If, after disconnecting a cubicle, a COM port other than the expected one disappears from the list, it may be that cables are swapped between the ports in the USB Hub and the corresponding cubicles.

Arena Settings: Define the Arenas

Create and Calibrate the Arenas

1. Choose Setup > Arena Settings.
2. Grab a background image from the cameras, calibrate, and draw the arenas. For details, see Arena Settings in the EthoVision XT Help.

   Tip: To quickly calibrate, draw lines between opposite walls of the cage, next to the grid, and enter 17 cm (the cage width).

3. If the image of a cage is not centered, see Camera Orientation for guidance.
4. Proceed with declaring the FCS devices for each arena.

Arena Settings: Declare the FCS Devices

Follow this procedure to specify the FCS devices that you want to use for each cage (arena).

1. In the Arena Settings, click the Arena – Hardware Mapping button at the bottom-right corner of the EthoVision XT screen.
2. The Arena - Device Mapping window opens. This window lists under Device groups the FC systems connected to the COM ports currently declared. The second column, Arena, lists the arenas currently associated with each system.
3. Check carefully that the name of each arena matches the physical FC system and the COM port associated with it. If they do not match, select the correct arena name under Arena.
4. Once you are ready, click OK. The Arena-Hardware Mapping window opens. This window lists the names of the actual devices that EthoVision XT can control. By default, each device is uniquely associated with an arena.

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