Mar 26, 2026
Morris Water Maze
- Aroa S Maroto1,
- Egoa Ugarte Perez1,
- Antonio Cerdán Cerdá1,
- Silvia De Santis1
- 1Instituto de Neurociencias (Consejo Superior de Investigaciones Científicas - Universidad Miguel Hernández), San Juan de Alicante 03550, Spain
Protocol Citation: Aroa S Maroto, Egoa Ugarte Perez, Antonio Cerdán Cerdá, Silvia De Santis 2026. Morris Water Maze. protocols.io https://dx.doi.org/10.17504/protocols.io.e6nvwn832vmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: January 30, 2026
Last Modified: March 26, 2026
Protocol Integer ID: 241862
Keywords: Congnitve test, water maze, aeging, morris water maze, memory in rodent, morris water maze the morris water maze, detecting cognitive alteration, assessing spatial learning, using distal spatial cue, distal spatial cue, cognitive alteration, spatial learning, swim path, rodent, measures such as escape latency, behavioural test, circular pool, used behavioural test, memory, animal
Abstract
The Morris Water Maze is a widely used behavioural test for assessing spatial learning and memory in rodents. In this protocol, animals are trained to locate a hidden platform in a circular pool by using distal spatial cues. Performance is evaluated using measures such as escape latency, swim path and time spent in the target quadrant. This method provides a reliable and sensitive way of detecting cognitive alterations associated with physiological and pathological changes.
Guidelines
This protocol was carried out using Wistar rats.
It is important to habituate the rats to the operator so that they can try to dismiss the street for the manipulation.
Materials
- Pool with a diameter of 180 cm and a depth of 60 cm
- Water maze automated Atlantis platform (Panlab) LE820300 760026
- Water heater for pool AM14-457
- Computer
- Blackfly S BFS-U3-16S2C camera, FLIR
- SpinView software for controlling camera settings
- Bonsai software for live animal tracking and recording
- Posters with different geometric shapes in black and white for use as cues.
- No toxic ink for the water.
- Heating lamp
- Towels
Troubleshooting
Safety warnings
- Maintain the water temperature between 19 and 22°C throughout the experiment.
- Adjusting the camera settings to avoid having reflections on the water will make reviewing the videos easier.
- Ensure that the waiting and action times remain constant to avoid introducing variability to your test.
Ethics statement
All experimental procedures were approved by the Animal Care and Use Committee of the Instituto de Neurociencias de Alicante (Spain) and were conducted in compliance with Spanish (Law 32/2007) and European regulations (EU Directive 86/609, EU Decree 2001-486, and EU Recommendation 2007/526/EC).
Before start
- Place all the cues.
- Fill the pool, leaving a 10 cm gap at the edge.
- Fix the platform 1.5 cm below the water level.
- Heat the water to 22°C.
- Check the camera focus and light.
- Add the dye to the water to make it dark.
- Prepare the drying box with the heat lamp and towel. The heat lamp should not occupy the entire box so that the animal can choose whether or not to stay in the warm area and avoid burns.
- The operator should leave the water maze area while the animal is in the pool.
Pre-setting
20m
Check camera settings
5m
Introduce the Atlantis platform into the pool and ensure it moves correctly, avoiding making waves during the up-and-down movements.
5m
Place the platorm in a diferent space from the previous day.
5m
The first day of the experiment is an exception to this rule.
Start the Bonsai workflow,
Check that it is recording at 30 fps.
5m
Trial 1
36m 20s
Put the platorm down , make it not possible to use for the animal
1m
Place the animal in the desired start position in the maze, facing the tank center.
1m
Do not throw the animal into the water. Lower it gently to avoid splashing.
30s
The moment the animal is released and the operator leaves the experimental area, a timer or computer tracking program is started.
After 1 minute, press the button to raise the platform.
Stop the timer when the animal reaches (touches) the platform A trial limit of 2 min.
2m
PlatformFrom 9 to 12 steps
Rats will find the platform. They will do this in two minutes or less
Return to remove the animal and leaving the cage for driying.
1m
Leave the animal for 30 minutes until start trial 2
30m
Trail 2-4
10m 20s
Put the platorm up, make it possible to use for the animal but not visible
1m
Place the animal in the desired start position in the maze, facing the tank center. Each trial should be started from a different position, chosen randomly.
1m
Do not throw the animal into the water. Lower it gently to avoid splashing.
30s
The moment the animal is released and the operator leaves the experimental area, a timer or computer tracking program is started.
Stop the timer when the animal reaches (touches) the platform or when is past 1 min
1m
Platform2 steps
Rats will find the platform. They will do this in 1 minute or less
Trial2-4
6m
Return to remove the animal and leaving the cage for driying.
1m
Leave the animal for 5 minutes until the next trial
5m
Protocol references
- Spooner, R.I.W. & Thomson, A & Hall, Jeremy & Morris, Richard & Salter, Stephen. (1994). The Atlantis platform: A new design and further developments of Buresova's on-demand platform for the water maze. Learning & memory (Cold Spring Harbor, N.Y.). 1. 203-11. 10.1101/lm.1.3.203.
- Vorhees, C., Williams, M. Morris water maze: procedures for assessing spatial and related forms of learning and memory. Nat Protoc 1, 848–858 (2006). https://doi.org/10.1038/nprot.2006.116
Acknowledgements
The authors gratefully acknowledge the professional staff of the Animal Facility (SEA) of the Universidad Miguel Hernández (UMH) for their excellent animal care.