Jan 15, 2025

Public workspaceOpen Field

DOI
dx.doi.org/10.17504/protocols.io.kqdg3qb2zv25/v1
  • 1University of Alabama at Birmingham
Ashley Adamson
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DOI: dx.doi.org/10.17504/protocols.io.kqdg3qb2zv25/v1
Protocol Citation: Ashley Adamson, Allie Smith, Briana R De Miranda 2025. Open Field. protocols.io https://dx.doi.org/10.17504/protocols.io.kqdg3qb2zv25/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 14, 2025
Last Modified: January 15, 2025
Protocol Integer ID: 118268
Funders Acknowledgements:
Briana De Miranda
Grant ID: R01ES034846
Briana De Miranda
Grant ID: TX220241
Ashley Adamson
Grant ID: F31 ES036890
Abstract
The following protocol is used to assess gross motor and anxiety-related behaviors using the Open Field Test at the University of Alabama at Birmingham's Neuroscience Behavioral Assessment Core.
Guidelines
Use of this protocol needs approval by user's Institutional Animal Care and Use Committee (IACUC) or equivalent ethics committee.
Materials
  1. Chlorhexidine
  2. Paper towels
  3. Open field boxes (appropriately sized for test subject)
  4. Ethovision version 17 software
Before start
You will need a test mouse whose behavior is not included in the experiment to calibrate detection settings.
Set Up
Set Up
Bring animals into testing room for habituation (at least 30 minutes prior to start of experiment)
Set up Maze
Set up the number of open field boxes you want to use (6 fit well under our camera setup)
Clean the maze
  • spray with chlorhexidine and wipe out
Plug the Ethovision version 17 software dongle into the computer
  • shines a red light when working
Open EthoVision version 17 software
Start a new experiment
Name the file
Indicate where the file will be saved
Experiment Settings
Select live tracking
  • select the camera to be used
  • center your arena(s) under the camera
Select center point detection
Select the number of arenas you will be using
Units should be cm, s, and deg
Arena Settings
Grab a background image from the camera
  • ensure the maze is in its intended area, moving it after this step will affect analysis
  • mark placement with tape so the maze can be returned to the exact same location if accidentally moved between trials
Draw scale
  • measure the width of one box
  • draw a scale bar over the area measured and indicates its length (in cm)
Click the first arena in the list on the right
  • select the subdivided rectangle drawing tool
  • trace one box
  • divide the rectangle into 4 rows and 4 columns
  • drag the arena label inside the box
  • repeat for all arenas
Create cumulative zones
  • Inside - add the 4 boxes in the middle of the subdivided rectangle to this cumulative zone
  • Outside - add the 12 boxes around the edge of the subdivided rectangle to this cumulative zone
Validate setup
  • If you have any overlapping zones, they will be considered invalid and must be redrawn
Trial Control Settings
Begin condition should be on the start button with the condition of center point being in the arena
Stop track with the condition being after 5 minutes
Detection Settings
Click advanced settings
Dynamic response
Indicate if subject is darker or lighter than background
Place a test subject in the maze
Click and drag to create a box around the subject
The software will create an automatic threshold around the subject that moves with it
Finetune it with the scrub bar to make it fit perfectly around the subject
Trial list
Add the desired number of trials
Running the Experiment
Running the Experiment
Acquisition
Software will load the first trial
Put the subject(s) in starting arena(s)
  • in the center of the arena
  • keep starting area consistent across subjects
Press record
  • the program will run for 5 minutes and automatically stop
Remove subject(s)
Clean enclosure with chlorhexidine
Repeat for all trials
Results and Analysis
Results and Analysis
Data profile
Ensure that the software is pulling data from all trials
Analysis Profile
Distance traveled
  • Arena
  • Statistic - total only
Velocity
  • Arena
  • Averaging interval - 1
  • Statistics - mean
Acceleration
  • Arena
  • Statistics - mean
In zone
  • Select the inside and outside cumulative zones
  • Frequency
  • Latency to first
  • Cumulative duration
Results
Statistics and charts
Calculate to generate a file with the analysis parameters chosen
Export and save
Ensure the video files are exported and saved
  • analysis can be repeated from video files if a problem occurs
Acknowledgements
We would like to thank the UAB Neuroscience Behavioral Assessment Core for use of their facilities.