Project protocol - Contents

• Workflow and sampling
• Equipment and supplies
• Reagents and solutions
• Procedure: Open field test
• Data

Workflow and sampling

Step	Procedure	Equipment	Data collected
1	Acclimation in test room (1h)	-	-
2	Open field test, over 3 days of testing	Open field arena	Locomotor activity and exploratory behavior

Equipment and supplies

• Open field chamber (Digiscan 16-beam automated activity monitor)
  • 42 x 42 x 30 cm
  • Within sound attenuating cubicle measuring 65 x 55 x 55 cm (Med Associates)
  • No lights used (testing in complete darkness)
• PC computer and Digiscan software
• Ventilating fan in ceiling of test room
  • 55 dB masking background noise

Reagents and solutions

• 20% isopropyl alcohol (for cleaning chamber)

Procedure: Open field test

1. Activity measures are conducted for 5 min on 3 consecutive days (tested between 1100 and 1700h) in the same order (order of testing various strains is randomized).
2. Approximately 1h prior to testing, mice are placed in the testing room for acclimation.
3. After acclimation, mice are transferred to a holding cage for 5 min before being placed in the center of a cleaned darkened chamber.
4. Following testing, mice are transferred to home cages and returned to the colony room.

Data collected by investigator

• Horizontal activity
• Horizontal bouts
• Time spent moving
• Time spent resting
• Vertical activity
• Vertical bouts
• Time spent vertical
• Stereotypic activity
• Stereotypic bouts
• Time spent moving stereotypically
• Clockwise revolutions
• Counter-clockwise revolutions
• Distance traveled
• Distance traveled in periphery
• Time spent in periphery
• Distance traveled in center
• Time spent in center
• Time spent in left-front of arena
• Time spent in right-front of arena
• Time spent in left-rear of arena
• Time spent in right-rear of arena

Percentages computed by MPD