That is an interesting application.
I make several improvement. Now I can distinguish the first type (classic biconcave cell with a hole in the middle) and the second type. I do not know which is the third type here, so I just ignore it.
A trick to distinguish these two type is to calculate the intensity at the center. Please see attached pipeline. Using classfyobject, you can find the difference between each cells.
RescaleIntensity. The signal is bit weak and background is high. This module makes image easy to distugish
Threshold. Make it binary.
Identfy the object
Shrink the obj_cell to a point and expand to 5-10 pixel large.
Measure the intensity within the object in 4.
If you have a lot of images and you want to distinguish the percentage of each type, please use this pipeline and export the result to .mat. Then, it is easy to use MATLAB to calculate the result.
(Cellprofiler works well with MATLAB)
Alphonse.zip (163.9 KB)