Knowledge Base

Trial Control Settings

The Trial Control Settings in EthoVision XT allows you to program activation of calcium imaging and optogenetic stimulation by Inscopix devices at specific times or when specific events occur, for example when the subject enters a target zone of the arena.

In addition, in Trial Control Settings you can define the conditions for the start and stop of data acquisition.

In EthoVision XT, choose Setup > Trial Control Settings > New, and enter a name for the new settings profile.

For More Information

For more information on how to work with Conditions and Actions in Trial Control Settings, see the EthoVision XT Help (press F1 in EthoVision XT) and the EthoVision XT 18 - Trial and Hardware Control - Reference Manual, which you can find in the Apps screen under Noldus > EthoVision XT 18 Other Documentation.

Example 1 - Image Recording (nVista)

Use Case

EthoVision XT starts tracking after locating the subject's center point in the arena. Next, EthoVision XT sends out a high TTL pulse to the TRIG port of nVista. After a time delay of 10 minutes, EthoVision XT stops tracking the subject.

Hardware Connections

See the scheme on page 343.

Prerequisites in EthoVision XT

You specified the TTL port and output line (either 1 or 2) for TRIG in the Experiment Settings and mapped the port to the arena (see page 346).

Procedure

1. In the Trial Control Settings, under Actions, click the button next to Custom hardware.
2. Specify the device (e.g. Device A) and the type of signal (Output 1 High).
3. Optionally, edit the name in the Action name field.
4. Click OK and insert the Action box between the Condition - In zone box and the Action - Start track box.
5. Click Settings in the Condition - Time box and select 10 minutes. This way tracking stops after 10 minutes.
6. If you want to stop recording in nVista when you stop behavior tracking, bring a new Action box in the flow line which instructs the system to send out a low TTL pulse to nVista. Make sure that you place the Action box to the right of the 10-minutes time box.

Tip: In the Hardware Action box you can test whether EthoVision XT triggers the nVista. To do so, click the Test button.

Note: Make sure to instruct nVista/nVoke to record video based on the TRIG signal.

Example 2 - Manual Video Recording, Triggered OG-LED Stimulation

Use Case

EthoVision XT starts tracking after locating the subject's center point in the arena. You start video recording in nVoke manually by clicking the Record button. At the planned time, for example at 10 minutes after start, EthoVision XT sends out a high TTL pulse to the GPIO[n] port of nVoke.

In the Inscopix DAQ software, under Video Recording configuration, make sure that Trigger is set to NONE and Control is set to Video Recording ON.

Hardware Connections

See the scheme on page 344. You do not need to use the TRIG port of the nVoke device.

Set the GPIO[n] to Input in the Inscopix DAQ software.

Prerequisites in EthoVision XT

In the Experiment Settings, you defined the TTL port and output line (either 1 or 2) connected to the GPIO[n] of nVoke. In Arena Settings, you mapped that port to the arena (see page 346).

Procedure

Because calcium imaging is started manually in this example, you only have to program the activation of the OG-LED. In the following example, the Time condition is set to wait 10 minutes before the triggering action.

1. In the Trial Control Settings, under Conditions, click the button next to Time.
2. Specify the time (e.g. 10 minutes). Optionally, edit the name in the Condition name field.
3. Click OK and insert the Condition box after the Action - Start track box (or wherever it applies in your protocol; for example in a sub-rule).
4. In the Trial Control Settings, under Actions, click the button next to Custom hardware.
5. Specify the device (e.g. Device A) and the type of signal (Output 1 High).
6. Optionally, edit the name in the Action name field.
7. Click OK and insert the Action box to the right of the Condition box just created.
8. When applicable, define a Condition and an Action (Output Low) for stopping the trigger action and insert them in the trial control flow line.

Notes

• In the Inscopix DAQ software, make sure to set the Trigger mode for the GPIO[n] port to Follow.
• When the Trigger is set to low, LED stimulation is stopped. To set the Trigger to Low, add an Action to the same device/TTL port and output line as above, and select the signal Output Low.
• You can also let the DAQ software run its own pulse protocol while the Trigger is ON. In that case the Control for the GPIO[n] port is set to Pulse protocol.
• For more information, see the nVista and nVoke User Manual.

Example 3 - Triggered Video Recording and LED Stimulation

Use Case

EthoVision XT starts tracking after locating the subject's center point in the arena. Both video recording and the OG-LED are triggered by EthoVision XT when the corresponding conditions (time or event) are met.

Hardware Connections

See the scheme on page 344.

Set the GPIO[n] to Input in the Inscopix DAQ software.

Prerequisites in EthoVision XT

In the Experiment Settings, you defined two TTL lines from the USB-IO box: one connected to the TRIG and the other to the GPIO[n] port of nVoke. In Arena Settings, you mapped the ports/lines to the arena (see page 346).

Procedure

Define actions and conditions as explained for the previous examples.

• Condition to trigger video recording.
• Action to trigger video recording through the TRIG port.
• Condition to trigger OG-LED stimulation.
• Action to trigger the OG-LED stimulation through the GPIO[n] port.

When applicable, define additional conditions and actions to stop video recording and OG-LED stimulation.

Notes

• In the Inscopix DAQ software, make sure to set the Trigger mode for the TRIG port to Follow, and the Control is set to Video Recording ON. In an alternative configuration, you can set Control to Recording schedule to record video for a specific time.
• In the Inscopix DAQ software, make sure to set the Trigger mode for the GPIO[n] port to Follow. Select the type of Control you require (e.g. Pulse protocol).
• See Example Configuration #3 in the nVista and nVoke User Manual.

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