Objective To explore issues of the human tissue biobank, ranging from its establishment, collection and preservation of samples, quality control, management and application. Methods Development of standardized operational procedures, collection of such samples as fresh frozen tissues from the surgery, paraffin-embedded tissues, whole blood, serum, plasma, and cerebrospinal fluid. Specimens were classified and made aliquots according to different requirements, and then stored at room temperature, -80℃ refrigerator or in liquid nitrogen. Microsoft Access database system was used in the management of these specimens. Results From September 2004 to September 2008, a total of 11 872 samples from patients with benign and malignant diseases were collected and preserved. Among them, 4360 tubes of fresh frozen tissue samples from 2500 cases were provided within and beyond the province. These samples were also applied to DNA, RNA, protein extraction and tissue microarray, and immunohistochemistry-related research. Conclusion Human tissue biobank is highly useful in sharing human tissue resources effectively, as it can provide high quality specimens from benign and malignant diseases and normal control. In addition, it plays a very important role in exploring pathogenesis, developing new technologies for early detection of disease and new therapeutic strategies.