Jun 02, 2026
Preparation for behavioural tests in mice
- Amandine EVEN1,2,3,4,5,
- Sriparna Mukherjee1,2,3,5,
- Louis-Eric Trudeau1,2,3,5,6,4
- 1Department of pharmacology and physiology, Faculty of Medicine, Université de Montréal;
- 2Neural Signaling and Circuitry research group (SNC);
- 3Center for Interdisciplinary Research on the Brain and Learning (CIRCA);
- 4Institut Courtois d’innovation biomédicale;
- 5Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD, 20815;
- 6Department of Neuroscience, Faculty of Medicine, Université de Montréal
Protocol Citation: Amandine EVEN, Sriparna Mukherjee, Louis-Eric Trudeau 2026. Preparation for behavioural tests in mice. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygxj834l8j/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: March 30, 2026
Last Modified: June 02, 2026
Protocol Integer ID: 314080
Keywords: behavioral testing, animal handling, habituation, habituation procedures for mouse, consistent behavioral performance across animal, preparation for behavioural test, mice this protocol, behavioural test, habituation procedure, mice, guidelines for experimenter blinding, consistent behavioral performance, experimenter blinding, mouse, habituation period, test order
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: ASAP-000525
Abstract
This protocol describes general considerations and habituation procedures for mouse behavioral testing.
It includes guidelines for experimenter blinding, group allocation, test order, timing, and environmental control to minimize variability.
It also details handling and habituation period to reduce stress and ensure consistent behavioral performance across animals.
General considerations
The experimenter should be blind to the treatment and genotype of the mice.
If the experimenter is working with a large group of mice, he/she/they must ask someone to divide the mice in three or more groups, making sure that each group has an almost equal number of control and treated mice for each genotype.
The sequence of tests should be the same amongst the groups. [If for group 1, the experimenter follows a sequence of open field, rotarod and pole test, then for group 2, he/she/they must follow the same sequence of tests.]
It is advisable not to perform two extensive tasks on the mice on the same day. [for example, open filed and rotarod should be done on two separate days]. Do not perform long experiments lasting full days, otherwise, the animals tested in the morning and afternoon will be from different phases of their light cycle. Try to complete your experiment in under 4 hours per batch.
Try to follow the same working hours to begin and end the behaviour tasks during the period of the experiment.
The experimenter should plug in and start the apparatus needed before moving the mice in the manipulation room as (1) it is little hard to locate things under red light, (2) the rotarod apparatus makes some noise when it is turned on.
On days when you plan to handle or test mice, do not use any perfume, body lotion, after-shave etc. to avoid stressing the mice.
Experiments should preferably be performed in an inverted light-dark cycle. It is preferable to start the behaviour work at least one hour after the light-dark cycle change. Avoid working with the mice exactly after the light-dark transition.
Habituation before each set of behavioral experiment
5d 0h 10m
The experimenter should handle the mice once a day for at least 5 days before beginning any behavioral experiments. This allows the mice to become familiar with him/her/them, and the effect of stress on mouse behavior will be minimized.
5d
The experimenter should spend almost equal time (at least 00:10:00 per mouse) with each mouse in the group during each day of handling.
10m