STED proteins images - setting conditions with WEKA?

Hi all!

I am new to imageJ and I would like to process some STED images of membrane proteins (see attached files).N1_rat_VRAC on PDL_2_B_sted_decon.tif (2.0 MB)

I was thinking of doing WEKA segmentation + additional thresholding and, eventually, using the “Analyze Particles” tools to detect/measure the shape and the density of the particles.

However, I am having trouble in choosing the best WEKA setting conditions because I don’t know how to avoid bias. I am checking the recall/precision plot as a measure of quality of segmentation, but I’m not sure is the best way. It’s not that trivial because the signal pattern is very complex and reminds much more of a “spray”, rather than well defined and large blobs to be distinguished from a clear background (like in cell counting).

Any suggestions?

Thank you in advance!