Your images very likely have different scaling for pixel and image size, or have scaling on some channel(s) and not on some other(s). Because of this, EzColocalization can not perform analysis. It might be worth double-checking your images to make sure they have the same field of view, time points, and magnification.
There are a couple easy fixes for your analysis. First, you could apply a global sizing to your images by selecting "Analyze" from the menu bar of ImageJ/Fiji, then "Set Scale..." from the drop down menu, clicking the "Global" check box, clicking the "OK" button, and then perform your analysis with EzColocalization. Second, if one of your images is lacking scaling, or has the wrong scaling for some image(s) you can remove the scaling from your images by selecting each of your images, selecting "Analyze" from the menu bar of ImageJ/Fiji, then "Set Scale..." from the drop down menu, clicking the "Click to Remove Scale" button, selecting the "OK" button for all your images, and then performing analysis as normal.
Your images very likely have different scaling for pixel and image size, or have scaling on some channel(s) and not on some other(s). Because of this, EzColocalization can not perform analysis. It might be worth double-checking your images to make sure they have the same field of view, time points, and magnification.
There are a couple easy fixes for your analysis. First, you could apply a global sizing to your images by selecting "Analyze" from the menu bar of ImageJ/Fiji, then "Set Scale..." from the drop down menu, clicking the "Global" check box, clicking the "OK" button, and then perform your analysis with EzColocalization. Second, if one of your images is lacking scaling, or has the wrong scaling for some image(s) you can remove the scaling from your images by selecting each of your images, selecting "Analyze" from the menu bar of ImageJ/Fiji, then "Set Scale..." from the drop down menu, clicking the "Click to Remove Scale" button, selecting the "OK" button for all your images, and then performing analysis as normal.