Tracking of migrating cells under phase-contrast video microscopy with combined mean-shift processes.
In this paper, we propose a combination of mean-shift-based tracking processes to establish migrating cell trajectories through in vitro phase-contrast video microscopy. After a recapitulation on how the mean-shift algorithm permits efficient object tracking we describe the proposed extension and apply it to the in vitro cell tracking problem. In this application, the cells are unmarked (i.e., no fluorescent probe is used) and are observed under classical phase-contrast microscopy. By introducing an adaptive combination of several kernels, we address several problems such as variations in size and shape of the tracked objects (e.g., those occurring in the case of cell membrane extensions), the presence of incomplete (or noncontrasted) object boundaries, partially overlapping objects and object splitting (in the case of cell divisions or mitoses). Comparing the tracking results automatically obtained to those generated manually by a human expert, we tested the stability of the different algorithm parameters and their effects on the tracking results. We also show how the method is resistant to a decrease in image resolution and accidental defocusing (which may occur during long experiments, e.g., dozens of hours). Finally, we applied our methodology on cancer cell tracking and showed that cytochalasin-D significantly inhibits cell motility.
Debeir
O
O
Department of Logical and Numerical Systems, Faculty of Applied Sciences, CP 165/57, Université Libre de Bruxelles, 50 Av. F. Roosevelt, 1050 Brussels, Belgium. odebeir@ulb.ac.be
Van Ham
P
P
Kiss
R
R
Decaestecker
C
C
D000230:Adenocarcinoma / Q000473:pathology* / Q000503:physiopathology; D000465:Algorithms; D001185:Artificial Intelligence; D045744:Cell Line, Tumor; D002465:Cell Movement; D003196:Computer Graphics; D003198:Computer Simulation; D006801:Humans; D007089:Image Enhancement / Q000379:methods; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008175:Lung Neoplasms / Q000473:pathology* / Q000503:physiopathology; D008856:Microscopy, Fluorescence / Q000379:methods*; D008858:Microscopy, Phase-Contrast / Q000379:methods*; D018715:Microscopy, Video / Q000379:methods*; D008954:Models, Biological; D009716:Numerical Analysis, Computer-Assisted; D015588:Observer Variation; D010363:Pattern Recognition, Automated / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012815:Signal Processing, Computer-Assisted; D013382:Subtraction Technique; D014584:User-Computer Interface