Home›Cell Biology›Quantitative analysis of Wound Healing, Cell Migration or Scratch Assay using ImageJ Software
Steps
1Download and install the wound healing macro00:07
2Open and convert image to 8-bit format02:06
3Configure macro parameters and set threshold02:28
4Define measurement properties and analysis settings03:43
5Execute macro and measure wound area04:37
6Edit region properties and customize visualization05:34
7Save results and export measurement data07:26
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Quantitative analysis of Wound Healing, Cell Migration or Scratch Assay using ImageJ Software
Protocol
Difficulty
intermediate
Steps
1
Download and install the wound healing macro
Download the MRI wound healing tool macro file (with .ijm extension) from the instructional website and install it into ImageJ through the Plugins > Macro > Install menu.
▶ 00:07
2
Open and convert image to 8-bit format
Open the scratch assay image in ImageJ software and convert it to 8-bit format if the original image is in RGB color format.
▶ 02:06
3
Configure macro parameters and set threshold
Right-click on macro option M to open the dialog box and set parameters including method (variance), variance filter radius (20), threshold value (35), and minimum area size for measurement.
▶ 02:28
4
Define measurement properties and analysis settings
Access Analyze > Analyze Particles to set size units (1 to Infinity in square inches), keep circularity settings, and enable Display Results and Overlay options.
▶ 03:43
5
Execute macro and measure wound area
Left-click on macro option M to run the analysis and display the scratch assay area with yellow boundary lines and measurement results including area and standard deviation values.
▶ 04:37
6
Edit region properties and customize visualization
Use the ROI Manager to adjust line properties such as color (change from yellow to red), thickness, and labels; uncheck label options to remove numerical displays for publication purposes.
▶ 05:34
7
Save results and export measurement data
Save the measurement results to an Excel file and save the analyzed image with overlay lines by selecting Save as Overlay in the ROI Manager dialog.