Hi Everyone,
I stained FFPE tissue with two different colors for two different antigens. I can deconvolute the original picture in FIJI (ImageJ), assign pseudofluorescent colors to the two deconvoluted channels, and merge them back together again to make one pseudofluorescent picture with two different colors.
However, I cannot find out how to assign a third color for the spots in which the two stainings are colocalized. I tried to use the coloc2 extension, but this produces only quantifications of colocalized area and no images to the best of my knowledge. Can anybody help me or maybe advise an alternative program for this purpose?
Cheers,
Jasper
Make each channel a binary mask, take the overlap of the two binary masks, use that itself to mask the original non-binary combined image. You should be left with a masked out area of the original image with full dynamic range, but only in the areas where they are colocalized. Then pseudocolor to whatever you'd like.
Hi,
Additional ideas and discussion on relevant colocalization issues:
Article Quantifying Receptor Trafficking and Colocalization with Con...
https://uu.diva-portal.org/smash/get/diva2:563664/FULLTEXT02.pdf
https://sydney.edu.au/medicine/bosch/facilities/advanced-microscopy/user-support/ImageJ_FL_Image_Analysis.pdf
https://cdn.auckland.ac.nz/assets/fmhs/sms/biru/docs/Colocalisation_Analysis_Tools_in_ImageJ.pdf
https://www.nature.com/articles/s41598-018-33592-8
http://www4.utsouthwestern.edu/livecellimagingfacility/docs/Phelps_Colocalization%20Chapter_MethMolBiol.pdf
Article Quantitative analysis of the factors that affect the determi...
I hope this helps :)
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