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This thesis mainly focuses on proposing a 4D (three spatial dimensions plus time) tissue-volume preserving non-rigid image registration algorithm for pulmonary 4D computed tomography (4DCT) data sets to provide relevant information for radiation therapy and to estimate pulmonary ventilation. The sum of squared tissue volume difference (SSTVD) similarity cost takes into account the CT intensity changes of spatially corresponding voxels, which is caused by variations of the fraction of tissue within voxels throughout the respiratory cycle. The proposed 4D SSTVD registration scheme considers the entire dynamic 4D data set simultaneously, using both spatial and temporal information. We employed a uniform 4D cubic B-spline parametrization of the transform and a temporally extended linear elasticity regularization of deformation field to ensure temporal smoothness and thus biological plausibility of estimated deformation. A multi-resolution multi-grid registration framework was used with a limited-memory Broyden Fletcher Goldfarb Shanno (LBFGS) optimizer for rapid convergence rate, robustness against local minima and limited memory consumption. The algorithm was prototyped in Matlab and then fully implemented in C++ in Elastix package based on the Insight Segmentation and Registration Toolkit (ITK). We conducted experiments on 2D+t synthetic images to demonstrate the effectiveness of the proposed method. The 4D SSTVD algorithm was also tested on clinical pulmonary 4DCT data sets in comparison with existing 3D pairwise SSTVD algorithm and 4D sum of squared difference (SSD) algorithm. The mean landmark error and mean landmark irregularity were calculated based on manually annotated landmarks on publicly available 4DCT data sets to evaluate the accuracy and temporal smoothness of the registration results. A 4D landmarking software tool was also designed and implemented in Java as an ImageJ plug-in to help facilitate the landmark labeling process in 4DCT data sets.