FIJI (ImageJ): Analysis of Scratch Assays, Wound Healing & Cell Migration [Wound Healing Size Tool]

Learn how to use FIJI (ImageJ) to analyze images of scratch wound healing or transwell migration assays. The plugin used (Wound Healing Size Tool) automatically recognizes wound healing size, corrects the average wound width by considering its inclination, and quantifies parameters such as: area, wound area fraction, average wound width, and width deviation of the wound images. It can be used for individual images (2D) or stacks (time-lapse).

Timeline:
00:00 Introduction
00:10 Single image (2D) and Wound healing size tool
02:17 Brightfield/Phase contrast stack analysis with Wound healing size Stacks Tool
04:25 Fluorescence image stack & Wound healing size Stacks Tool
05:49 Wound healing size Manual tool

Plugin reference: Suarez-Arnedo, A., Torres Figueroa, F., Clavijo, C., Arbeláez, P., Cruz, J. C., & Muñoz-Camargo, C. (2020). An image J plugin for the high throughput image analysis of in vitro scratch wound healing assays. PloS one, 15(7), e0232565.

[job]

Hi Johanna, it's the usual amazing you here again! Thank you extremely well. Please, is there a way I can save the image files with the main trace lines retained? Best!

[perrikaty]

Thank you for your video! I followed your instructions and it worked for my bright field images. However, no matter what parameter I put in, it didn't work for my fluorescence images as it appeared "Index (0) is outside of the 0—1 range in line 284 \ roiManager (“select”, 0 <)> ;". Could you please let me know how to fix this? Thanks!

[ravichandrans]

Hey, Thank you soo much for the clarification. ❤❤....I have a doubt, What value I can use to calculate the wound closure percentage. I don't know how to create a graph with these values....Kindly help me.

[jessyhell]

Oi Johanna, tudo bem?


O meu problema é que a extensão do Wound healing size não está instalando. Sempre que tento de acordo com a descrição do seu vídeo, aparece a mensagem: "There are no images open in line 5
Var width = [getWidth];

[tremmble]

Hello Johanna, Thank you for the video! I have one error that I am dealing with (Error: Index (0) is outside of the 0--1 range) which I think occurs when the wound closes completely. Is there a way I can still output a results table with a 0 for the area fraction for all images in a sequence where the wound is closed?

[khinaye]

How can I change the unit of width from pixel to micrometers

[pattarapoungcho]

Hello thank you for nice video and helpful. I would like to download wound healing tool. But I visited that site you posted I can’t find that plug in on zip file. Only have pdf file. Thank you

[edagultekin]

Hello, thank you for this video. I am running this code in my batch process to apply it to multiple images. How can I obtain a list of complete measurements with labels? I receive single results one by one. THank younin advance.

[smitakapoor]

Hello Johanna I have one query while following all the steps as mentioned by you but the area of wound is not getting selected rather it is showing some other selected region than the scratch region what might be the reason. kindly resolve this. Thank you in advance.

[rushilshanbhog]

Really helpful video. I was wondering if you don't have a time-lapse stack file and just two jpg files, is there any way to use the stacking tool in imageJ to see the relation between area and time?

[NatashaAley]

Thank you for this! When I use my images I get several ROI's appear but only 1 measurement. How do I know which ROI matches to the measurement?

[job]

In addition, once I re-open ImageJ software where will I find the installed Wound Healing Size Tool plugin?

[comfortwilliams]

Dr. Thank you for this video. They are indeed helpful for my image analysis.
I appreciate you.