Computational optical-sectioning microscopy for 3D quantification of cell motion: results and challenges

How cells move and navigate within a 3D tissue mass is of central importance in such diverse problems as embryonic development, wound healing and metastasis. This locomotion can now be visualized and quantified using computational optical-sectioning microscopy, which permits non-destructive 3D imaging of living specimens over long time periods. This technique, however, presents several technical challenges. Image restoration methods must be fast enough to process numerous I Gbyte time-lapse data sets (16 Mbytes per 3D image×60 time points). Because some cells are weakly labeled and background intensity is often high due to unincorporated dye, the SNR in some of these images is poor. Also required are accurate, automated-tracking procedures to generate both 3D trajectories for individual cells and 3D flows for a group of cells. Finally, sophisticated visualization techniques are needed to view the 3D movies of cell locomotion. Here, I discuss our current approaches to these problems and note present limitations