i am trying to design a pipeline for several weeks without success. My main problem is i am not able to outline the cells.
I have a set of images of cells that are staining for 3 different components: DAPI (blue), Prot1 (green immunostaining), Prot2 (red immunostaining).
I have a set images of control cells and a few sets of cells with treatments.
What we want to do is measure the effect of those treaments on the level of Prot2 (red).
Prot1 is our reference signal; it covers the whole cell.
Prot2 give a cortical signal, mostly on the edge of the cells.
Can you suggest a simple pipeline?
What i have tried so far:
It seems i can’t outline the cells properly.
Then, when i do measurements of intensity, the results are the same for control cells and positive controls, on which the level of Prot2 should be a lot decreased.
Can you give me any advice on threshold settings, etc.?
I have uploaded a set of 3 pics (DAPI, Prot1, Prot2).
Any help would be appreciated!!!
Controls.zip (220 KB)