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The Barnes Maze, a powerful tool in behavioral neuroscience, serves as an essential apparatus for studying spatial learning and memory in rodents. This comprehensive guide unveils the fundamental components and methodology required to effectively utilize this maze for insightful research outcomes.

Key Components of the Barnes Maze

The Barnes Maze is a circular maze designed to assess spatial learning and memory in rodents. Its dimensions and the materials needed to complete the assay are as follows:

Barnes Maze Dimensions:

  • Maze Diameter: The Barnes Maze typically has a diameter ranging from approximately 88 to 120 centimeters about 35 (mice) to 47 (rats) inches. The larger the maze, the more challenging it can be for rodents to find the target.
  • Circular Platform: The maze consists of a circular, flat platform elevated above the ground. The platform is typically made from non-slip materials to provide stable footing for rodents.
  • Multiple Holes: Around the perimeter of the platform, evenly spaced holes are present, usually numbering between 2 to 20 holes. These holes are escape holes that lead to the maze’s underside.
  • Escape Box: Beneath one of the holes, there is an escape box where the rodent can find refuge. The escape box provides a safe and secure location for the rodent to rest and escape from the maze.
  • Visual Cues: Visual cues are placed around the maze’s perimeter. These cues serve as spatial reference points to help rodents orient themselves and navigate.

Materials Needed for the Barnes Maze Assay:

  • Barnes Maze: Acquire a Barnes Maze constructed from durable materials such as acrylic or plastic. Ensure the maze’s dimensions conform to standard specifications.
  • Escape Box: Prepare escape boxes that fit snugly beneath the escape holes. These boxes should be made from a material that provides comfort and security for the rodents.
  • Visual Cues: Set up visual cues around the maze’s perimeter. These cues can be images, symbols, or objects that serve as reference points for the rodents.
  • Start Location: Designate a starting point on the maze, often marked by a designated zone or platform where the rodent begins each trial.
  • Video Cameras: Install video cameras to record the rodent’s movements and behavior within the maze. Multiple camera angles may be needed to capture comprehensive data.
  • Video Recording Equipment: Set up video recording equipment, including cameras, lighting, and a recording system, to capture and store behavioral data during the experiments.
  • Cleaning Supplies: Have cleaning supplies on hand to maintain the cleanliness of the maze between trials, ensuring the removal of any residual odors that could affect rodent behavior.
  • (Optional) Control Software: Utilize appropriate control software to manage the experiment, including tracking the rodent’s path, controlling escape holes, and recording data.
  • (Optional) Data Analysis Software: After the experiment, use data analysis software to analyze the collected behavioral data and assess spatial learning and memory performance.

These materials, when used in accordance with the specified dimensions and setup described, create an effective platform for conducting Barnes Maze experiments to investigate spatial cognition and memory in rodents. Researchers can adapt specific details based on their experimental requirements and objectives.

Preparing the Maze

Before commencing experiments, ensure the Barnes Maze is clean, free from lingering odors, and devoid of any disturbances that might affect rodent behavior. Place the maze in a controlled environment with consistent lighting, temperature, and minimal noise to maintain data reliability.

Animal Selection and Habituation

Carefully select rodents that align with your research objectives, considering factors like age, gender, and genetic background. Allow them to acclimate to the testing room to reduce stress and anxiety levels. Gradual habituation to the maze environment is vital to minimize initial apprehension and novelty-induced stress.

Conducting Spatial Learning Trials

Initiate the experiment by placing the rodent at the designated starting point and allowing it to explore the maze. Researchers often employ the following steps for spatial learning trials:

  1. Training Phase: During this phase, the rodent learns to locate the escape box. Researchers record the time taken to find the escape box and the number of errors made.
  2. Probe Trial: After several training trials, a probe trial is conducted without the escape box to assess memory retention. Researchers monitor the time spent in the target quadrant where the escape box was previously located.

Behavioral Parameters Assessed by the Barnes Maze

The Barnes Maze is a versatile tool for studying spatial learning and memory in rodents, allowing researchers to assess various behavioral parameters, including:

  1. Latency to Find Escape Box: The time it takes for the rodent to locate and enter the escape box. Longer latencies may indicate impaired spatial learning.
  2. Number of Errors: The number of incorrect choices or visits to non-escape holes before finding the escape box. Increased errors suggest difficulties in spatial memory and learning.
  3. Path Length: The total distance traveled by the rodent while searching for the escape box. Longer path lengths may indicate inefficiency in navigating the maze.
  4. Thigmotaxis: The tendency of the rodent to remain close to the maze’s perimeter instead of exploring the central area. Increased thigmotaxis can suggest anxiety-related behaviors.
  5. Time in Target Quadrant: During probe trials, the amount of time spent in the quadrant where the escape box was previously located. This parameter assesses memory retention.
  6. Probe Trial Errors: The number of incorrect choices or visits to non-escape holes during probe trials, where the escape box is absent. Increased errors can indicate deficits in memory recall.
  7. Revisits: The number of revisits to the same holes or locations previously explored. Excessive revisits may suggest inefficient spatial memory.
  8. Velocity: The speed at which the rodent navigates the maze. Changes in velocity can reflect alterations in motor function or motivation.
  9. Strategy Used: Researchers can categorize the strategies employed by rodents to locate the escape box, such as serial searching, random exploration, or systematic searching.
  10. Spatial Preference: Assessing whether the rodent displays a preference for particular escape hole locations or specific paths within the maze.
  11. Adaptation: Changes in behavior across trials, reflecting the rodent’s ability to adapt and improve its performance over time.
  12. Learning Curves: Graphing and analyzing the rodent’s performance over multiple trials to visualize the rate of learning and memory improvement.
  13. Stress-Related Behaviors: Observing stress-related behaviors such as freezing, grooming, or defecation during maze navigation, which can provide insights into emotional responses.
  14. Anxiety-Related Behaviors: Examining anxiety-related behaviors, such as thigmotaxis or hesitancy to explore open areas, which may indicate anxiety or fear responses.
  15. Cognitive Flexibility: Assessing the rodent’s ability to switch strategies when the location of the escape box changes between trials.

By analyzing these behavioral parameters in Barnes Maze experiments, researchers can gain valuable insights into spatial learning and memory, as well as the effects of various interventions or treatments on cognitive performance. This data can be instrumental in understanding and addressing cognitive dysfunction, memory disorders, and neurological conditions in preclinical research.

Data Analysis

Post-experiment, analyze the collected behavioral data to evaluate the rodent’s performance in terms of spatial learning and memory. Interpret the results within the context of your research objectives to gain insights into various aspects of rodent cognition.

Barnes Maze Data Interpretation & Relevance of Pharmaceutical Intervention

The behavioral parameters assessed in Barnes Maze experiments hold significant relevance to pharmaceutical interventions for drug treatments of cognitive dysfunction and dementia. Here’s how these parameters are important in the context of drug development and treatment:

  1. Latency to Find Escape Box: Reduced latency suggests improved spatial learning, which is a crucial cognitive function. Pharmaceutical interventions that lead to shorter latencies may indicate enhanced memory and learning abilities in individuals with cognitive dysfunction or dementia.
  2. Number of Errors: A decrease in errors signifies better memory recall and spatial learning. Drugs targeting cognitive dysfunction aim to reduce errors, as this reflects improved cognitive function.
  3. Path Length: Smaller path lengths suggest more efficient navigation, which is indicative of improved cognitive processing. Pharmaceutical interventions should ideally lead to shorter path lengths, demonstrating enhanced cognitive abilities.
  4. Thigmotaxis: A decrease in thigmotaxis indicates reduced anxiety-related behaviors. Some individuals with cognitive dysfunction or dementia may exhibit increased anxiety, and drugs that alleviate thigmotaxis may improve overall well-being.
  5. Time in Target Quadrant: During probe trials, increased time in the target quadrant signifies better memory retention. Drugs that improve memory function should lead to longer stays in the target quadrant.
  6. Probe Trial Errors: Reduced errors during probe trials indicate enhanced memory recall and retention. Pharmaceutical interventions aim to decrease errors in memory-related tasks.
  7. Revisits: A decrease in revisits suggests more efficient spatial memory. Drugs that reduce revisits may enhance the individual’s ability to remember explored locations.
  8. Velocity: Changes in velocity may indicate alterations in motor function or motivation. Understanding these changes can help assess the impact of pharmaceutical interventions on cognitive and motor aspects of dementia.
  9. Spatial Preference: Changes in spatial preference can reflect alterations in memory and spatial cognition. Drugs that restore or modify spatial preference may improve cognitive function.
  10. Learning Curves: Analyzing learning curves provides insights into the rate of cognitive improvement. Pharmaceutical interventions that steepen learning curves may accelerate cognitive recovery.
  11. Stress-Related Behaviors: Reducing stress-related behaviors is important for overall well-being. Some pharmaceutical interventions may aim to decrease stress-related responses, which can be beneficial for individuals with cognitive dysfunction or dementia.
  12. Anxiety-Related Behaviors: Addressing anxiety-related behaviors is essential for improving the quality of life for individuals with dementia. Drugs that mitigate anxiety-related responses can enhance overall comfort.
  13. Cognitive Flexibility: Assessing cognitive flexibility helps determine whether drugs can enhance adaptability in individuals with cognitive dysfunction. Cognitive flexibility is essential for daily tasks and problem-solving.

By evaluating these behavioral parameters in Barnes Maze experiments, researchers can gain valuable insights into the effects of pharmaceutical interventions on cognitive function, memory, and emotional well-being. This information guides the development and evaluation of potential treatments for cognitive dysfunction and dementia, ultimately aiming to improve the lives of affected individuals.


  1. Pitts M. W. (2018). Barnes Maze Procedure for Spatial Learning and Memory in MiceBio-protocol8(5), e2744. doi:10.21769/bioprotoc.2744.
  2. Van Den Herrewegen, Y., Denewet, L., Buckinx, A. et al. The Barnes Maze Task Reveals Specific Impairment of Spatial Learning Strategy in the Intrahippocampal Kainic Acid Model for Temporal Lobe Epilepsy. Neurochem Res (2019) 44: 600.
  3. Gawel, K., Gibula, E., Marszalek-Grabska, M., Filarowska, J., & Kotlinska, J. H. (2019). Assessment of spatial learning and memory in the Barnes maze task in rodents-methodological considerationNaunyn-Schmiedeberg’s archives of pharmacology392(1), 1–18. doi:10.1007/s00210-018-1589-y.

Additional information

Weight 50 lbs
Dimensions 40 × 6 × 40 in

Mouse, Rat


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