Related Experiment Video
Updated: Feb 13, 2026
Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice
Published on: February 6, 2015
[Measuring anxiety with behavior analysis software and comparing human observations with EthoVision XT in the
Introduction:
The reproduction of anxiety in laboratory animals is a renewing problem whenever a new drug is to be tested for its anxiolytic effect. Some gold-standard tests, such as the elevated plus maze test, are always considered to be used as reference. However, many controversial results for different anxiolytics were reported in elevated plus maze test. The analysis methods used by different labs could be the source of variability of the results. Human observations were the most commonly used since the 90's, when behavior analysis software appeared. In each lab, specific procedures for reducing bias in ethopharmacological experiments were implemented, but the performance of human observers was rarely compared to software assisted analysis.
Methods:
Four analysts and 24 trials were involved, each analyst having to do all the analyses during which they had to register eight parameters. All trials were also analyzed with the EthoVision XT (Noldus IT, Netherlands).
Results:
Several crucial parameters of the elevated plus maze test were significantly different between the analysts (p<0.05). The results registered by human observers were summarized and compared to the results of automatic analysis, which showed significant difference in the case of closed arm entry and total distance (p<0.001).
Conclusions:
In this study, it was shown that despite all precautionary measures taken to reduce the variability and bias among observers the results were clearly different from those registered by behavior analysis software. As a conclusion, it can be stated that the behavior analysis methods need some kind of standardization in order to be comparable between labs, preferably the use of the same software and/or settings.
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