Automated Scratch Assay Analysis - doesn't work?

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Automated Scratch Assay Analysis - doesn't work?

imagenoob
Hi guys,

I'm hoping to get imageJ to automatically measure the scratch performed on the scratch assay. I have downloaded the MRI Wound Healing Tool, but while it worked on sample images, if doesn't work on images created in our lab:




Does anyone have any tips? I've already tried playing around with the contrast, subtracting background, using Gaussian Blur, to no effect.

Many thanks for your time!
CL
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Re: Automated Scratch Assay Analysis - doesn't work?

Gabriel Landini
On Wednesday, 22 February 2017 17:26:50 GMT imagenoob wrote:
> I'm hoping to get imageJ to automatically measure the scratch performed on
> the scratch assay. I have downloaded the MRI Wound Healing Tool, but while
> it worked on sample images, if doesn't work on images created in our lab:
>
> <http://imagej.1557.x6.nabble.com/file/n5018174/scratch_test.png>
> <http://imagej.1557.x6.nabble.com/file/n5018174/scratch_test_imagej.png>
>
> Does anyone have any tips? I've already tried playing around with the
> contrast, subtracting background, using Gaussian Blur, to no effect.

You need to ask somebody who knows how to set up the phase contrast microscope
properly so there is an even background illumination. Your current setup is
incorrect and no amount of imaging will compensate for that.

Compare your images with some random image in the net with the microscope
properly set up:
http://jcb.rupress.org/content/jcb/suppl/2008/01/14/jcb.200708194.DC1/1.html

Cheers

Gabriel

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Re: Automated Scratch Assay Analysis - doesn't work?

Rhodri Wilson
In reply to this post by imagenoob
Gabriel is correct the microscope isn't set up correctly, I may have something to help but will have to check if it can be distributed. In the meantime why don't you look into shading correction to help with your image artifacts.

Rhod

Dr Rhodri Wilson
Microscopy Image Analyst
Microscopy Core
Department of Oncology
Old Road Campus Research Building
Old Road Campus
Roosevelt Drive
Oxford
OX3 7DQ

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Re: Automated Scratch Assay Analysis - doesn't work?

Rhodri Wilson
In reply to this post by imagenoob
If you contact me via a real email I cant share with you our scratch would analysis plugin.
Rhod

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