Immunofluorescence is widely used in detection and characterization of circulating tumor cells (CTCs). The process of immunofluorescence staining on cells needs centrifugal force to wash residual fluorescence, which is not only time consuming but also cells harmful. To speed up the staining process, a three-dimensional (3D) microfluidic device is employed in this work. This study also reports the rapid selection of identified live CTCs from whole blood on SACA chip for further characterization by automatic imaging and in-situ cell capturing process. The whole process takes 120 minutes for examining 4 ml whole blood sample and the recognition sensitivity can approach 1/108 by using the integrated chip and automatic image system. All cells are remained highly viable after the rapid selection process for further applications.