Wound Healing Help



I am trying to analyse some images from a wound healing assay. The example pipeline and it is good at finding the “tissue” in my control (T0.tiff), but not so good at identifying the cells once they have started to migrate (T24.tiff/T48.tiff/T72.tiff). As these cells move quite independently (rather than holding hands!) perhaps I need to adjust the settings to account for this?

Any advice/tips much appreciated!

Please find my images here: https://drive.google.com/drive/folders/0B9WqsdNBlJLELWppX1BvM1Nxc2s?usp=sharing

Many thanks



I guess it’s more meaningful for you to quantify the area covered by cells (which is increasing as the wound is being healed), rather than focus on segmentation of every single cells, isn’t it?

If so, you may try to increase the contrast of the cellular area vs. the empty background. For example by using Smooth, or using some series ImageMath as I made here wound_heal.cpproj (642 KB)
Here I ignore the coarse segmentation of single cells, but focus on identifying the area covered by cells.

Hope your research heal every wounds !


This is great! Many thanks for your help!


Hello again!

I am trying to run some additional images through this pipeline, but am not having much success- can you advise how I should tinker with the pipeline to improve this?

Please find my images here https://drive.google.com/open?id=0B9WqsdNBlJLERGc2WTl1QVlCQzA

Many thanks in advance!


Can you explain what “not having much success” means? It’s easier to help if you can point to a more specific issue. Thanks!


Hi Beth,

Sorry, I should have been more clear!

The pipeline isn’t distinguishing the cell(/wound) outlines:



The images in this second set had much more variable light conditions than the first set you posted, which is probably a big part of why the pipeline did poorly the second time. You might want to try something based on using the Enhance Texture model; I’ve posted a version that needs to be tweaked (it can fail when the cells are really confluent) but should give you a road map of a direction to take.

Good luck!

wound_heal_enhance.cppipe (9 KB)