Knowledge Base

Data Analysis

Simple Data Selection

Choose Analysis > Data Profile > New.

To Select the Track Segments When the LED Was Activated

1. Under Nesting choose Hardware state.
2. From the Device type list, choose Custom Hardware.
3. Choose the Device, the Signal (Is output 1 High), Value (true).
4. Click OK and insert the Nest box in the Data profile sequence.
5. Choose Analysis > Analysis Profile > New and define the variables you want to calculate for when the animal was stimulated (for example, the distance moved).

To Select the Track Segments When the LED Was Not Activated

1. Under Nesting choose Hardware state.
2. From the Device type list, choose Custom Hardware.
3. Choose the Device, the Signal (Is output 1 High), Value (false).
4. Click OK and insert the Nest box in the Data profile sequence.
5. Choose Analysis > Analysis Profile > New and define the variables you want to calculate for when the animal was not stimulated.

Advanced Data Selection

To compare two sets of track segments (LED activated vs LED not activated), create the two Nest boxes as described above. Place each box in a separate branch ending with a Result box. To create another Result box, under Common Elements choose Result.

Note that if you use the Pulser to generate high frequency optical pulses, the individual pulses are not "seen" by EthoVision XT. Because EthoVision XT controls the Pulser via the single TTL start/stop commands ("Output 1 High/Low"), it only "sees" the time that the command was sent out (Output 1 High/Low). Therefore, the pulse sequence is seen as a one time segment.

For more information on data selection, see the EthoVision XT Help.

To Select the Track Segments When a Particular Frequency Was Generated (Setup with Pulser)

Prerequisite: your Trial Control procedure includes control of frequency of stimulation. Two Subrules are defined: one for zone A (stimulation at 25 Hz) and one for zone B (stimulation at 50 Hz). Within a Subrule, Hardware actions "Activate Pulser" and "Deactivate Pulser" can be used to mark the time that a particular frequency was used.

1. In the Data Profile, under Nesting choose Trial Control State.
2. Choose the name of the actions that refer to a particular frequency:
   • From Element Action: [action that triggers the Pulser]
   • To Element Action: [action that stops the trigger]
   From the Event lists, select becomes active for both elements.
3. Click OK and insert the Nest box in the Data profile sequence.
4. Choose Analysis > Analysis Profile > New and define the variables you want to calculate.

Calculating Statistics

Once you have selected the track segments you are interested in, choose the variables you want to calculate. Choose Analysis > Analysis Profile > New.

To calculate the number of times that the optical stimulation was activated, in the Analysis profile choose Hardware command. Choose the following:

• Device type = Custom Hardware
• Device = the LED controller (or Pulser)
• Command = Output 1 High
• Under Trial Statistics choose Frequency

To calculate the total time that stimulation was activated, in the Analysis profile choose Hardware state. Choose the following:

• Device type = Custom Hardware
• Device = the LED controller (or Pulser)
• Signal = Is Output 1 High
• Value = true
• Under Trial Statistics choose Cumulative Duration

To calculate the statistics, choose Analysis > Results > Statistics and Charts.

Integrated Visualization

Choose Analysis > Results > Integrated Visualization.

The Trial Control protocol uses stimulation of 25 Hz when the animal is in zone A and 50 Hz when the animal is in zone B. The following variables can be displayed:

• In zone. Shows when the animal was in zones A and B. To show this variable, you must define it in the Analysis profile (choose In zone under Location).
• Hardware command. Marks the time when the Output 1 High was sent out from EthoVision XT.
• Hardware state. Marks the time interval when Output 1 stayed High.
• Trial Control state - Pulser 25 / Trial Control state - Pulser 50. Marks the time that the Hardware command within one of the Subrules was High. That command triggered the pulser for a specific stimulation (25 or 50 Hz). This way instances of Hardware state can be split according to the frequency of the stimulation.

Installing the Prizmatix Pulser

Follow this section if the Prizmatix Pulser/PulserPlus is not properly installed on the EthoVision XT computer.

Software Installation

Insert the Prizmatix Pulser software CD in the EthoVision XT computer's CD/DVD ROM drive, or download the most recent version from www.prizmatix.com/software.htm under Pulser/PulserPlus Software.

Double-click setup.exe. Follow the instructions on the screen and at the end of installation restart the computer. Follow the instructions below to update the driver.

Note: The installation procedure differs depending on whether you have the Pulser or PulserPlus. For more information, see the Prizmatix Pulser/PulserPlus User Manual.

Driver Update

Do this for each Pulser you use, when they are not recognized.

1. Connect the Pulser to the computer with the supplied USB cable.
2. In the Control Panel, open the Device Manager.
3. Under Other Devices, right-click Unknown device and select Update driver.
4. Choose Browse my computer for driver software.
5. Click Browse and select the following folder: C:\Program Files (x86)\Prizmatix Pulser [version number]\Drivers, then click Next.
   Note: This location may differ on your PC if you downloaded the drivers and saved them to another location.
6. In the Windows Security dialog click Install.
7. At the end of the update process click Close.
8. In the Device Manager, under Ports, you should see Prizmatix DRV.

To configure the Pulser driver for EthoVision XT, see the configuration section.

Calcium Imaging

• Introduction
• Physical Setup
• Basic Settings
• Program Control of Devices in EthoVision XT
• Detection Settings
• Aligning Data
• Data Analysis in EthoVision XT

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