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Koehler et al developed the Visual Stimuli Box to evaluate mouse vision through looming visual stimuli. The setup includes a square enclosure with an opaque shelter, a ceiling-mounted computer monitor for projecting stimuli, and a camera for recording behavior (monitor and camera optional).

The looming stimulus is an expanding black circle on the monitor. Mice are placed in the enclosure to acclimate before being exposed to the stimulus, and their responses (flight or freeze) are recorded and analyzed with motion-tracking software. This apparatus is compatible for tracking and analysis using video tracking packages such as ConductVision, Noldus EthoVision, or ANY-Maze

This device is a simple, reliable tool for assessing vision-related behavioral responses, focusing on image-forming pathways in the central visual system.

Price & Dimensions

Mouse + monitor

$ 4300

Per Month
  • Chamber: 40 cm L × 50 cm W × 30 cm H
  • Includes software for visual stimuli
  • Includes monitor

Mouse + monitor +video tracking

$ 6300

Per Month
  • Chamber: 40 cm L × 50 cm W × 30 cm H
  • Includes software for visual stimuli
  • Includes monitor
  • Includes 1-year licence for ConductVision

Rat + monitor

$ 5300

Per Month
  • Chamber: 50 cm L × 65 cm W × 40 cm H
  • Includes software for visual stimuli
  • Includes monitor

Rat + monitor + video tracking

$ 7300

Per Month
  • Chamber: 50 cm L × 65 cm W × 40 cm H
  • Includes software for visual stimuli
  • Includes monitor
  • Includes 1-year licence for ConductVision

Need tracking software?

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Introduction

The Looming Visual Stimuli box was originally designed to assess rodent visual acuity and response to looming stimuli. For the experiment, mice were acclimated in the enclosure for 10 minutes to explore and familiarize themselves with the environment. Once acclimated, the looming stimulus was presented when the mouse was away from the shelter. Responses such as freezing or fleeing to the shelter were recorded.

Motion-tracking software analyzed behavioral changes, including velocity and proximity to the shelter. The experiment reliably elicited fear responses in healthy mice, demonstrating its effectiveness in testing image-forming vision pathways. Blind mice showed no response, validating the vision-dependent nature of the test.

This simple, robust setup is a cost-effective tool for assessing visual behavior and studying neural pathways associated with vision.

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Apparatus and Equipment

The Looming Visual Stimulus assay includes the following components:

  1. Enclosure (provided): A rectangular open-lid enclosure (40 × 50 × 33 cm) constructed from acrylicThe floor is lined with paper for easy cleanup between trials. An opaque shelter is placed in one corner, with the entrance facing the center of the arena to facilitate easy access for the mice.
  2. Computer Monitor (optional): A 21-inch monitor mounted on the ceiling of the enclosure to project the looming visual stimuli. A secondary monitor duplicates the images for external observation.
  3. Visual Stimulus (software included): The looming stimulus consists of an expanding black circle programmed using MatLab and PsychToolbox3. It starts at a visual angle of 2° and expands to 50° over 250 milliseconds, repeating 10 times with a 1-second interval.
  4. Camera (optional)
  5. Motion-Tracking Software (optional): Used to analyze the recorded video and quantify changes in velocity and distance from the shelter, providing insights into the mouse’s behavioral responses to the stimuli. Our software, ConductVision can be used for this task.

Training Protocol

1. Preparation for the Experiment

1.1 Prepare the rectangular open-lid enclosure (40 × 50 × 33 cm).
1.2 Place an opaque shelter in one corner with the entrance facing the center of the arena.
1.3 Mount a computer monitor on the ceiling to display stimuli and use a secondary monitor to duplicate the display.
1.4 Set up a camera with a wide-angle lens adjacent to the enclosure to record mouse behavior. Use a frame rate of 60 FPS or higher.
1.5 Program a looming stimulus (expanding black circle) using our included software. Start at 2° visual angle and expand to 50° over 250 milliseconds. Repeat the stimulus 10 times with 1-second intervals.

2. Mouse Acclimation

2.1 Place a mouse in the enclosure and allow it to explore freely for 10 minutes to acclimate to the environment.
2.2 Drop food pellets in the corner opposite the shelter to encourage exploration.
2.3 Minimize stress during transfer by using the back of your hand as a resting place.

3. Looming Visual Stimuli Projection

3.1 Prepare the stimulus software before placing the mouse in the enclosure to minimize disruptions.
3.2 Start video recording 10 seconds before the stimulus begins.
3.3 Initiate the looming stimulus when the mouse is away from the shelter. If the mouse is unwilling to explore far corners, project the stimulus when moving freely elsewhere.
3.4 Present the stimulus 10 times, waiting 10 seconds after the last presentation before stopping the recording.

4. Mouse Transfer and Cleanup

4.1 Return the mouse to its original cage.
4.2 Clean the enclosure with soap and water.
4.3 Reposition the shelter to its initial location if it was moved.

5. Video Analysis

5.1 Use motion-tracking software to analyze mouse behavior before, during, and after stimulus presentation.
5.2 Calculate changes in velocity and distance to the shelter to classify behaviors as freezing (speed <20 mm/s for 0.5 s) or flight (speed >400 mm/s leading to shelter).

Data Analysis

The following parameters can be observed on the Looming Visual Box:

  • Flight response
  • Flight velocity
  • Freeze response 
  • Length of freeze

Summary

  • The Visual Water Box is used to study learning and memory behaviors in rodents when presented with a visual stimulus.
  • The Visual Water Box consists of a trapezoidal-shaped pool that is partially divided on one end creating two arms. At the end of each arm, a screen is placed, which displays visual stimuli.
  • Visual capabilities of rodents can easily be measured using the Visual Water Box and can be done with great speed.
  • The Visual Water Box can also be used to test the effects of diseases and disorders, brain lesions, and pharmacological manipulations on learning and memory.

References

Koehler CC, Hall LM, Hellmer CB, Ichinose T. Using Looming Visual Stimuli to Evaluate Mouse Vision. J Vis Exp. 2019 Jun 13;(148):10.3791/59766. doi: 10.3791/59766. PMID: 31259889; PMCID: PMC6686198.