Fig. 2

Visualization of processing steps in ROF filter based segmentation. a — Original single-channel image (ROI No. 09, CD14 ⁺/488 nm), contrast enhanced by factor 3, scale bar 45 μm. b — Cartoon of a as result of Steps 1 and 2, contrast enhanced by factor 6. c — Features to be examinated in b after masking with initial threshold i=3 (Steps 4 − 6). d — Feature classification in c (Steps 7 − 9): saved by Rule 1 for further processing (yellow); excluded by Rule 2 (red), Rule 3a (purple) or Rule 3b (pink); accepted as macrophages (green). Rule 3c caused no exclusions here. e — Features to be examinated in b after masking with incremented threshold i=4 (white); pixels saved in d but masked now (grey) (Step 10). f — Feature classification in e, color encoding as before. Rule 3c caused no exclusions again