How to Measure Anxiety in Mice & Rats on an Elevated Plus Maze

The elevated plus maze is a piece of equipment and protocol used by behavioral scientists to measure anxiety levels in laboratory rodents. Different-sized mazes can be used for either rats or mice, but the same principles apply to each experiment. The plus maze has four arms, arranged in a + shape; two of these arms have sides, so providing some shelter and a more secure feeling for the animal, and the other two arms are open. The premise behind the elevated plus maze is that more anxious rodents will spend less time in the uncovered, exposed arms of the maze and less anxious rodents will explore the open arms more freely.

Things You'll Need

  • Experimental behavior room
  • Elevated plus maze
  • Camera
  • Computer
  • Tracking software
  • Experimental mice/rats
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Instructions

  1. Setup

    • 1

      Arrange the elevated plus maze in a behavioral testing room directly underneath the ceiling-mounted camera.

    • 2

      Connect your computer to the camera and make sure that the image is clear --- you will need to see where the animal is within the maze at all times of testing.

    • 3

      Design a protocol on your tracking software. You will need to design a five-minute-long test for each animal and you will need to track its movements, recording how long the animal spends in each arm of the maze.

    • 4

      Place your experimental animals in the same room or an adjoining room for at least 30 minutes prior to testing to allow them to habituate.

    Testing

    • 5

      Place an experimental animal in the center of the plus maze, where all of the arms meet. Place the animal facing one of the open arms. The experiment begins as soon as the animal is placed on the maze.

    • 6

      Measure the amount of time spent in each arm using the tracking software. You may also wish to note and measure other behaviors such as rearing.

    • 7

      Remove the animal from the plus maze after the experimental time has elapsed and place it back in its home cage.

    • 8

      Clean the maze between each test with water.

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