Hello! My name is Diana, I am using immunohistochemistry images in which the protein of interest appears intense brown and cell nuclei have a purple color but are very dim:
Here is a zoom-in to the image in which the purple nuclei can be seen better
I need to normalize the number of protein plaques to the number of cells (identified by the number of nuclei). I can quite well count the protein plaques with IdentifyPrimaryObjects, I did not manage yet to count the nuclei. I have tested various modules:
-IdentifyPrimaryObjects module directly specifying nuclei size ranges - identifies only nuclei that are in the middle of protein plaques:
-Unmixcolors - trying to separate the purple color from the rest, did succeed at it
-EnhanceOrSupressFeatures - trying to enhance Dark Holes, since the region occupied by nuclei corresponds to “holes” within the protein-stained mesh, does not identify them correctly
-CorrectIlluminationCalculate - aiming to enhance the intensity of nuclei, did not succeed
Do you have any other ideas that could work?? Would it be possible to seize the fact that regions covered by nuclei correspond to “holes” in the protein mesh?
I would enormously appreciate any opinion!! Thank you so much in advance!