Dec 04, 2025
- Mario Alberto Bautista-Carro1
- 1Cinvestav
External link: https://doi.org/10.1371/journal.pone.0339028
Protocol Citation: Mario Alberto Bautista-Carro 2025. Open field test. protocols.io https://dx.doi.org/10.17504/protocols.io.yxmvmbqo5g3p/v1
Manuscript citation:
Bautista-Carro MA, Sánchez-Teoyotl P, Juárez-Serrano D, Iannitti T, Díaz A, Flores G, Morales-Medina JC (2026) Olfactory bulbectomy induces neurobiological alterations in the prefrontal cortex and hyperlocomotion in male rats. PLOS One 21(1). doi: 10.1371/journal.pone.0339028
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: October 07, 2025
Last Modified: December 04, 2025
Protocol Integer ID: 229260
Keywords: Locomotor activity, Exploration, Novel environment, Anxiety-like behavior, exploration in the open field test, open field test, open field test the open field test, test, field, open enclosure, laboratory, rodent, scientific research method, rat
Abstract
The open field test is a scientific research method that measures the general locomotion and willingness to explore of a rodent by placing it in a large, open enclosure. This protocol uses a black square arena without a top cover, in a field divided into 25 squares of equal size, with 300 LUX lighting. During the rat's exploration in the open field test, the researcher leaves the laboratory. To control for possible effects due to circadian cycles, all tests are performed at the same time of day. All tests are recorded on video with a central view of the field at a height of 2 m above the ground.
Guidelines
All tests should be conducted at approximately the same time of day. During the rat's exploration of the open field, the researcher should leave the laboratory.
Materials
Lux meter
Box with square base (85 x 85 cm) in black
Lamp
Canon video camera (model Vixia HF R70) with a 2.8-89.6 mm lens
Camera tripod
70% alcohol
Troubleshooting
Before start
Verify that there is uniform 300 LUX lighting within the open field and clean it with 70% alcohol on the base and walls. Transfer the rats to the behavior laboratory and wait 1 hour for habituation before beginning the open field test.
After 1 h of habituation in the laboratory, place the rat in the center of the field and allow it to explore for 5 min.
Remove the rat from the field and place it in a new box separate from its group.
After each trial, clean the base and walls of the open field test with 70% alcohol to eliminate olfactory cues.
Record locomotor activity as well as the frequency of rearing and grooming behaviors in the rats, using a blind observer.
Protocol references
Bautista-Carro, M. A., Flores, G., Iannitti, T., Galindo-Paredes, G., 26 Morales-Medina, J. C. (2021). Curcuma longa administration significantly reduces acute and persistent inflammatory pain measures in male and female rats. Arch. Vet. Sci. Med., 4(02). doi:10.26502/avsm.022
Morales-Medina, J. C., Dumont, Y., Benoit, C. E., Bastianetto, S., Flores, G., Fournier, A., 26 Quirion, R. (2012). Role of neuropeptide Y Y1 and Y2 receptors on behavioral despair in a rat model of depression with co-morbid anxiety. Neuropharmacology, 62(1), 200 208. https://doi.org/10.1016/j.neuropharm.2011.06.030