Four Dimensional Image Registration for Intravital Microscopy

Increasingly the behavior of living systems is being evaluated using intravital microscopy since it provides subcellular resolution of biological processes in an intact living organism. Intravital microscopy images are frequently confounded by motion resulting from animal respiration and heartbeat. In this paper we describe an image registration method capable of correcting motion artifacts in three dimensional fluorescence microscopy images collected over time. Our method uses 3D B-Spline non-rigid registration using a coarse-to-fine strategy to register stacks of images collected at different time intervals and 4D rigid registration to register 3D volumes over time. The results show that our proposed method has the ability of correcting global motion artifacts of sample tissues in four dimensional space, thereby revealing the motility of individual cells in the tissue.