I have stained a brain slice with antibodies to human nestin and human nuclear antigen. I’ve been able to identify the human nuclei (primary), and create a secondary (the cell) and tertiary object (cytoplasm). Using the secondary object and the grayscale nestin image, I was able to measure the intensity of nestin expression in my objects. I’ve also classified my cells based on that intensity.
I would like to be able to see if nestin expression changes the farther out a cell is from the initial tumor mass. Is there any method that can help me measure the individual distances of my objects (the cells) from a specified point (For the sample picture I am posting, I would like the point to be the far left-middle, which is about the center of the tumor mass)?
Also, while I’m posting this, for future use, is there a good way to identify objects that are more fiber/filament-like (Nestin)? I’ve been trying different threshold methods to identify nestin as objects, but it’s hard to match the staining.
I have attached a sample image and my pipeline so far. Any suggestions or feedback would be greatly appreciated!
IntensityClassification.cp (13.4 KB)