Scratch Assay analysis


I’m new to image J and trying to count the number of cells in my pictures for scratch analysis. I think I’ve managed to do it by doing: 8-bit image - image - adjust - threshold - apply - sharpen - find edges - analyse particles - summarise. However, when the number of cells should increase after day 0 (day of scratch) it goes down?

Does anyone know how to get around this please? The number of cells should increase as the scratch has covered over.

Would really appreciate any help.

do you really get a segmentation of the individual cells this way? What people often do is to measure the area of the gap over time. If you use the number of cells why do you need to do the ‘summarize’ step in your workflow? Do your images look similar to the images here:

Check what the objects you get after the analyze particles step are. I would suppose they are just random parts of the tissue and not individual cells.

If you send an image I could probably say more.

Best regards,

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if you like you can give this one a try:



As @volker requested… an example with original images would be best to share with us. So we can try to re-create the issue you are having in ImageJ. Please upload original image files and then the exact workflow you are currently using - then we can better help you.