Image stabilization and registration for tracking cells in the microvasculature

We propose a registration system to be used for tracking cells in intravital video microscopy that 1) stabilizes jitter-the undesired translational displacement of frames due to respiratory movement, etc ., and 2) registers frames in a moving field of view (FOV) to allow for cell tracking over an extended range. For the first time, tracking of rolling leukocytes in vivo over a moving FOV is demonstrated. In a fixed FOV, stable background regions are located using a morphological approach. Template subregions are then selected from the stable regions and matched to corresponding locations in a reference frame. We show the effectiveness of the stabilization algorithm by using an active contour to track 15 leukocytes previously untrackable due to jitter. For 30 fixed FOV sequences containing rolling leukocytes, the resulting root-mean-square error (RMSE) is less than 0.5 μm. To align frames in a moving FOV, we present a modified correlation approach to estimate the common region between two consecutive fixed FOVs. We correlate the overlapping regions of the initial frame of the current fixed FOV and the final frame of the previous fixed FOV to register the images in the adjoining moving FOV. The RMSE of our moving FOV registration technique was less than 0.6 μm. In 10 sequences from different venules, we were able to track 11 cells using an active contour approach over moving FOVs.