Can anyone point towards a tutorial or video where I can perform co-localization analysis using ImageJ? I have a very simple experiment where I have stained HEPG2 cells with DAPI in order to count the number of cells and I have transduced a vector expressing GFP. I really just need to know the number of cells that are green in a given image. What would be the best way to do this?
Hi Krishna Chintaluri . I don’t think you really need colocalization analysis based on your description. You just need to split the 2 channels first (Dapi and GFP), then threshold each channel. You will need certain tools in ImageJ to get your desired results: Thresholding, Analyze Particles, ROI Manager. Here are some tutorials to get you started:
FIJI (ImageJ): Counting 2D Objects
https://youtu.be/5b7x2qx1FF0
FIJI (ImageJ): Quantification of 2D Images (Measuring Area, Intensity, etc.)
https://youtu.be/XkGK7T12OiM
FIJI (ImageJ): Regions of Interest and the ROI Manager
https://youtu.be/kor9E3Qen28
FIJI (ImageJ): Measuring Overlap using the ROI Manager
https://youtu.be/4umlxVsjY04
Krishna Chintaluri Please check these options as :
https://youtu.be/rRyJnFo57xU
you can easily install there plugins form ImageJ website.
The easiest plugin I've found to use is called JaCoP (Just another colocalisation plugin)... Very easy to use :)
Hannah Bruce ,Chathura Abeywickrama , Johanna Marie Dela Cruz , thank you for your replies. I will be sure to try your recommendations.
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