Choose a template file that contains the number of chamber that you wish to use (e.g. 1, 2, or 4 arenas)
Arena Settings: Choose "Arena Settings" from the left sidebar, duplicate that last version of Arena Settings. Capture background video; this will open a live window with the overhead video. Adjust the chambers so they are captured by the camera in their entirety and are well-lit. Adjust the size, shape, and placement of the arenas, which are demarcated by oval shaded areas and a label, eg “Arena1” with an arrow pointing to the area of interest. Make sure no arena overlaps with any other, and that the arena for each chamber includes the entire bottom ring of the chamber, as well as some of the bottom side walls (for when an animal rears against the side wall).
Detection Settings: Choose "Detection Settings" from the left sidebar, duplicate the last version of Detection settings. Click on grab background image. Now place your mouse or mice into the chambers. You will see a live behavioral detection signal on screen. Where a mouse is detected, it will appear yellow, and its center point will be marked with a red dot. In the Detection Settings dialog box, you can adjust the contrast and the size detection parameters to optimize detection of the mouse. Slide these around until you reliably detect each mouse, but do not pick up reflections on the side wall. Sometimes the detection will vary in different parts of the arena, due to shadows or lighting conditions, so make sure the mouse moves to different areas of the arena before accepting the settings.
Trial Control Settings: Choose "Trial Control Settings" from the left sidebar. Choose the type of trial (eg “10 minute open field”) that you want. Testing sessions can be of variable length, but for basic characterization, 10-20 minutes is adequate.
Trials List: Choose "Trials List" from the left sidebar. If new empty trials will be needed, push the button at the top to add trials. Otherwise, enter the information for each mouse into the Trials List spreadsheet, and into your lab notebook. At minimum note the name of the Noldus Template file, the Trial #, Arena #, and mouse name, as well as the manipulation that day in your lab notebook.