Quantitative analysis of Wound Healing, Cell Migration or Scratch Assay using ImageJ Software

Link to the plugin:

[za]

Thank you👍🏻
What does it mean, change the threshold according the image type? How should I know what to choose for it?

[haileydeckers]

Hi, I’m new to Image J and not sure I’m adjusting the threshold etc correctly. When I press OK after the particle analyzer, it comes up with a box telling me “A threshold has not been set using the Image->Adjust->Threshold tool or the setThreshold(min, max) macro function. I think it is having trouble defining the edge, my setting are currently 30; 50; 4; 10000. Sorry

[KieraParker-fw]

Hello, great tutorial. You say at the end that you can incorporate your scale bar, how do you do this? Thanks!

[meganboseley]

Hello, How do I do this for a stack and save the images with the scratch area outline for all of them?

[ViquiiG]

I have the same problem as the others, I cannot define correctly the threshold. I've put the same as de image--> adjust --> threshold tool but it's still not working. I've tried putting the imagen in 16 bit and 1 threshold but did not work either.

[andrea-qwmy]

Hello, im new with ImageJ, when i wanted to install the plugin, it brings me into another page instead of downloading it. How can i fix it?

[SUPERANUJMAN]

Hey! Great informative tutorial mate, thanks. My requirement is to represent multiple outlines overlaid in a graph. Is there any method to that?

[drviolanwachukwunicholas-o]

Pls can you send me the protocol you used to induce scratch and type of microscope used to take images?

[pragatisingh]

Hi, thanks for this video. I have on issue with this method is at the last step when click on m, it calculates area of empty space that is wound but simultaneously it gives additional areas and yellow marks in between cells also. How I can remove/ avoid these areas which i dont want. Thanks!

[srishtichakraborty]

Hi. Great tutorial! I have a problem- I am unable to save the image as overlay in the last step. My image with the outline is not getting saved. Could you help me out with this?

[pegah_]

Hi thank you for your great video. I have a problem at the very beginning steps, i have cropped my circular picture taken from microscope, turned into square inserted into image j. Then 8bit-> find edges-> smooth
However when i click fill holes it fills the whole picture and if i dont do this after the scratch assay using steps from your video, it mistakenly circles around my cells instead of the scratch area. Could you please fell me what i should do

[bonicholas]

Hello, I'm new to ImageJ and not good with it yet! I followed all of the steps in the video, but when I left click on "m" at the end it just comes up with an ROI manager - do you know what I am doing wrong?

[njsantos]

Hello,
I installed the macro and followed all the steps. However, when I clicked "m", the "Wound healing tool Options" box did not appear. Where might I be going wrong? I use Windows. Thank you.

[saranyarajendran]

Hi thank you for this wonderful macros. I have the very similar problem as one of the user and however I change the values it is not working. Once I give values in analyze particle and click ok, it is giving message as 'A threshold has not been set using the Image->Adjust->Threshold tool or the set Threshold(min, max) macro function'. My parameters after clicking m button are as follows

i. Method: Variance
ii. Variance filer radius: 10
iii. Threshold: 20
iv. Radius close: 4
v. Min. Size: 5000

As you suggested for another user I tried changing the last value to triple digit numbers but nothing changed.
I am unable to measure these images. Please help! Thanks.

[drviolanwachukwunicholas-o]

Did you use 4x or 2x lens for taking scratch images?

[litipattanayak]

After doing the same..
I am getting the only m option, b is not coming... Can you please help me

[manichemist]

Is Set scale need to be filled? because i don't see any results after completing the procedure

[MarjanGhanbari-ht]

Hello, how can i set these for stack images ?

[sandhyap]

I don't find a m I.e method option in my toolbar. Wr do I find it ?