Cell capturing and separation is frequently involved in researches in immunology, diagnostics and pathology. In recent years, different new methods for cell capturing and separation based on microfluidic platform have been developed. With microfluidic chip, rapid and low cost detection can be implemented with small sample and reagent volumes. In this review, two methods of microfluidic cell separation, immune-capture and label-free separation, are introduced. Immuno-capture, a more traditional method, can separate cells with high specificity and provide highly purified target cells. Newly developed label-free method performs separation by utilizing the physical and biological features of target cells, and can keep cell's integrity and activity. Although there are a couple of issues to be resolved before microfluidic cell separation gets widespread commercial use, such as multi-system integration and industrial-scale manufacture, it has already attracted extraordinary research attention and has been in the path on rapid development. It is believed that cell capturing and separation methods based on microfluidic chip have broad application prospect in life science and biomedical field.