Knowledge Base

Arena Settings

Define the Arenas Automatically

Follow these instructions if your experiment was created for well plates with 6, 12, 24, 48 or 96 round wells, or 96 square wells, and this is the first new Arena Settings in your experiment. In other cases, see Define the Arenas Manually.

Tip: Use well plates with no notches between wells. These result in better well detection. Also make sure that the borders of all wells are within the camera image.

1. Place the well plate with water in the DanioVision chamber.
2. Choose Setup > Arena Settings > Arena Settings 1.
3. In the Grab Background Image window, click the Grab button to grab a background image of an empty well plate from the camera image.
4. If EthoVision XT can detect the wells, the outline of the arenas are displayed over the background image. A message appears asking whether the arenas overlap with the wells in the image.
5. Do one of the following:
   • If there is good overlap, click Yes. If needed, you can always make small adjustments using the Arrange Arenas function.
   • If the arenas clearly do not overlap with the wells, or arenas overlap with each other, click No. The Arrange Arenas window appears automatically. See Arrange the Arenas for details.
6. Click the 1. Draw Scale to Calibrate button. Draw a line over the entire well plate. Next, enter its size. Do this horizontally as well as vertically.
7. Click Validate Setup to validate the Arena Settings. Common errors are overlapping arenas, or zones falling outside an arena. If the setup is valid, continue with the instructions below.

Notes:

• Arenas are marked with labels A1, A2,... B1, B2,... according to your well plate (letters indicate rows; numbers indicate columns).
• Success of automatic detection depends greatly on image contrast, focus, and distortion. Therefore, automatic detection cannot be guaranteed to work in all situations or with video made with DanioVision versions earlier than DVOC-0040. Automatic detection also does not work if water is not present in the well plate.
• If automatic detection of the wells does not work, it could be due to the well plate not being centered in the video image, with some wells not visible or only partially visible. See Fine Camera Adjustments for guidance.
• When the well plate image is optimal and centered and EthoVision XT still shows the message "Well plate detection failed," click OK and proceed with Define the Arenas Manually.
• To start automatic well plate detection, click the Well Plate Detection button at the bottom of the Arenas and Zones section of the Arena Settings pane.

Define the Arenas Manually

The following instructions apply when the automatic well plate detection does not work properly. In that case you must define one arena and then make duplicates. If you used a predefined template to set up your experiment, go to Arrange the Arenas.

1. Click the 1. Draw Scale to Calibrate button, draw a line over the entire well plate and enter its size. Do this horizontally as well as vertically.
2. If the current arenas are of a shape different from that of the wells, select all arenas and press Delete.
3. Click 2. Select Shape and Draw Arena and use one of the shapes to create the first, top-left, arena. Make sure the label of the first arena (for example A1) points to the inside of the drawn arena.
4. Click the outline of the arena or its label and then click the 4. Copy Arena button. In the window that appears, select All Other Arenas, select the number of rows, and click OK.
5. Make sure the predefined arenas cover the wells. If you need to adjust the arena size and position, see Arrange the Arenas.

Arrange the Arenas

Use the Arrange Arenas function to move, re-scale, and rotate all arenas simultaneously.

1. In the Arena Settings window, click the 5. Arrange Arenas button to open the Arrange Arenas window.
2. To move all arenas, use the controls under 1. Move Grid:
   • To move up or down: Click the first slider and move it right (to move arenas down) or left (to move arenas up).
   • To move right or left: Click the second slider and move it right (to move arenas right) or left (to move arenas left).
   • You can also click the up/down arrow buttons of the corresponding box or enter a number in the box.
3. To scale all arenas, use the controls under 2. Scale Arenas:
   • Resize all shapes using the sliders, by pressing the up/down arrows of the corresponding box, or by entering a number in a box.
   • To increase the height of all arenas, use the first slider. To increase the width of all arenas, use the second slider.
   • By default, Keep Proportions is not selected, allowing you to resize height and width separately. If you select Keep Proportions, changing either the height or width will maintain the aspect ratio.
4. To rotate all arenas, use the controls under 3. Rotate Grid:
   • Optionally rotate all shapes. This is only necessary if the well plate is not positioned straight. The shapes rotate around the center of gravity of Arena 1.
5. To change the space between arenas, use the controls under 4. Arena Spacing:
   • By default, Keep Proportions is not selected, allowing you to change the space between shapes separately in the vertical and horizontal direction. If you select Keep Proportions, changing the spacing in either direction will maintain the aspect ratio.
6. Click Validate Setup to validate the Arena Settings. Common errors are overlapping arenas, or zones falling outside an arena. If the setup is valid, continue with the instructions below.

Tip: First position the upper-left arena in the desired position and set the desired size using Scale. Next, adjust arena spacing so all arenas are correctly positioned. See Using the Multiple Arena Setup in the EthoVision XT Help for more information on setting up multiple arenas.

Edit a Single Arena

Click the row in the Arena Settings pane for the arena you want to edit. The corresponding arena is highlighted. Click the handles to move or resize the polygon. For details on editing shapes, see Arena Settings > Work with Shapes in the EthoVision XT Help.

Assign the Stimuli to the Arenas

Use this procedure to apply stimuli during a trial, such as a light pulse or a tapping stimulus.

If you created an experiment with a pre-defined template and selected the correct DanioVision arena template, the White Light Stimulus and the Tapping Stimulus have been assigned automatically to the arenas and the steps below are not necessary.

Prerequisite: You have declared the port connections in the trial control hardware settings.

1. At the bottom of the Arena Settings pane, click the Arena - Hardware Mapping button.
2. In the Arena - Hardware Mapping window, do the following:
   1. Click Add Device.
   2. Locate the new row added to the list.
   3. Under Arena 1, select the device you want to add, for example White Light Stimulus or Tapping Stimulus.
   4. Repeat steps a through c for each additional device.
3. Click OK. Note that devices only need to be assigned to Arena 1, as they are applied to all arenas.

To test a device: Select the corresponding row in the Arena - Hardware Mapping window and click the Test button. See also Test the Hardware Devices in the EthoVision XT 18 Trial and Hardware Control Reference Manual.

For more information:

• About the Tapping Stimulus: see The DanioVision Tapping Device.
• About the TopLight Unit: see The DanioVision Toplight Unit.
• About the Optogenetics add-on: see The DanioVision Optogenetics Add-on.

Source: DanioVision DVOC-0050 Reference Manual - Arena Settings, Noldus Information Technology