Knowledge Base

Running the Spontaneous Behavior Protocol (EV11_01_SB3D / 02_SB3D2L)

1. This protocol is for experiments where the Day/Night cycle is 07:00-19:00 LIGHT; 19:00-07:00 DARK. If you are using a different Day/Night cycle these times need to be adjusted in the protocol. Please check with Sylics if this is the case for your experiments.
2. Before starting the experiment, mice should be single-housed for about a week. Going from socially housed in the holding rooms to single-housed on the day of testing can influence the behavior.
3. Prepare the PhenoTyper (bedding, water bottle, provide a tissue in the shelter as nesting material).
4. Introduce the mice to the PhenoTyper between 14:00 and 16:00 and place the top unit on the cage.
5. Start the protocol.
6. The protocol will run for 3 days (if protocol 1 is selected) or for 2.5 days (if protocol 2 is selected). If you are planning to run only the Spontaneous Behavior protocol, the 3 day protocol is recommended. If another protocol will run after the Spontaneous Behavior protocol (for example the CognitionWall protocol) the 2.5 days protocol is recommended.

Running the 1-Night CognitionWall Protocol (EV11_09_Wall1DFR5)

1. This protocol is for experiments where the Day/Night cycle is 07:00-19:00 LIGHT; 19:00-07:00 DARK. If you are using a different Day/Night cycle these times need to be adjusted in the protocol. Please check with Sylics if this is the case for your experiments.
2. Before starting the experiment, mice should be single-housed for about a week. Going from socially housed in the holding rooms to single-housed on the day of testing can influence the behavior.
3. Give the mice between 10 and 15 sucrose pellets on the day before testing (the same as they will have to earn during the test) so they get used to this unfamiliar food.
4. Prepare the PhenoTyper (bedding, water bottle, provide a tissue in the shelter as nesting material, remove food from the food tray or replace the food tray with a white PVC cover).
5. Fill the Pellet Dispensers with a small amount of pellets (maximum 1/3 of the container).
6. Weigh animals, introduce them into the PhenoTyper at 10:00 AM and place the top unit on the cage.
7. Between 16:00 and 16:30 (the start time of the CognitionWall protocol), lift the top unit and introduce the CognitionWall. The bedding material should not be inside the CognitionWall. Make sure the CognitionWall is in a stable position and the tube of the Pellet Dispenser protrudes all the way into the CognitionWall compartment.
8. Start the protocol.
9. The protocol will run for 24 hours (16:30), but can be manually stopped around 10:00.

Running the 4-Night CognitionWall Protocol (EV11_03_Wall4DFR5)

1. This protocol is for experiments where the Day/Night cycle is 07:00-19:00 LIGHT; 19:00-07:00 DARK. If you are using a different Day/Night cycle these times need to be adjusted in the protocol. Please check with Sylics if this is the case for your experiments.
2. Before starting the experiment, mice should be single-housed for about a week. Going from socially housed in the holding rooms to single-housed on the day of testing can influence the behavior.
3. Give the mice between 10 and 15 sucrose pellets on the day before testing (the same as they will have to earn during the test) so they get used to this unfamiliar food.
4. Prepare the PhenoTyper (bedding, water bottle, provide a tissue in the shelter as nesting material, remove food from the food tray or replace the food tray with a white PVC cover).
5. Fill the Pellet Dispensers with a small amount of pellets (maximum 1/3 of the container).
6. Weigh animals, introduce them into the PhenoTyper at 10:00 AM and place the top unit on the cage.
7. Between 16:00 and 16:30 (the start time of the CognitionWall protocol), lift the top unit and introduce the CognitionWall. The bedding material should not be inside the CognitionWall. Make sure the CognitionWall is in a stable position and the tube of the Pellet Dispenser protrudes all the way into the CognitionWall compartment.
8. Start the protocol.
9. The protocol will run for 96 hours (16:30), but can be manually stopped around 10:00 after the 4th night.

Zone Definitions

• Spontaneous Behavior protocols (SB3D and SB3D2L)
• CognitionWall protocols (1 & 2 night IL and 4 night IL and RL)
• Calibration of the PhenoTyper when using CognitionWall protocols

Source: EthoVision XT 18 - THC - Trial and Hardware Control, Noldus Information Technology