Jun 01, 2026
Openfield Test
- 1California Institute of Technology
Protocol Citation: Sayan Dutta 2026. Openfield Test . protocols.io https://dx.doi.org/10.17504/protocols.io.261geke7dg47/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: September 09, 2025
Last Modified: June 01, 2026
Protocol Integer ID: 226806
Keywords: openfield test, open field test, behavioral assay in rodent, behavioral assay, used behavioral assay, locomotor activity, rodent, anxiety, movement pattern, exploratory behavior
Abstract
The open field test is a widely used behavioral assay in rodents that evaluates locomotor activity, exploratory behavior, and anxiety-like responses in a novel environment. It measures parameters such as total distance traveled, movement patterns, and time spent in the center versus periphery of the arena.
Prior to the start of testing, animals were acclimated to the behavior room for a period of 30 minutes to minimize the influence of novel environmental stress on behavior.
After acclimation, animals were placed into a square open-field arena, which was subdivided to allow simultaneous monitoring of up to four individuals within the same session.
Each open-field test session lasted a total of 12 minutes. The first 2 minutes were designated as a habituation period, allowing the animals to adjust to the arena, while an overhead camera continuously recorded their behavior.
Locomotor activity and related summary statistics were quantified from the final 10 minutes of the recording using EthoVision XT 10 software (Noldus Information Technology).
At the end of each session, the arena was thoroughly cleaned to remove olfactory cues, with particular care taken when transitioning between species to avoid cross-contamination of scent or behavioral influences.