Medical device performance has been advanced while images are expected to be acquired with further higher quality and pertinent applicability as images have been increasing in importance in analyzing major organs. Recent high frequency of image processing by MATLAB in image analysis area accounts for the intent of this study to segment pulmonary vessels by means of MATLAB. This study is to consist of 3 phases including pulmonary region segmentation, pulmonary vessel segmentation and three dimensional connectivity assessment, in which vessel was segmented, using threshold level, from the pulmonary region segmented, vessel thickness was measured as two dimensional refining process and three dimensional connectivity was assessed as three dimensional refining process. It is expected that MATLAB-based image processing should contribute to diversity and reliability of medical image processing and that the study results may lay a foundation for chest CT images-related researches.