Background A biobank is a storage facility which stores sample mainly for biological or medical researches. Biospecimen collection is the kind of work essential in the building of biobank. However, we found the collection works had the difficulties in the proceeding because of the low anticipation rate. Methods We here conducted a questionnaire including several questions covered several aspects in order to find out people’s attitude towards biospecimen donation. The questionnaire had 20 questions mainly focused on overall publics’ participation rates, matters that influence their participation and major publics’ concerns in biospecimen donation. Results In our survey, 1477 from 2200 distributed questionnaires were responded, and electronic questionnaires showed the highest response rate of 49.9%. In all respondents, 936 showed willingness in participation, providing a percentage of 63.4%. We found that most respondents lack the knowledge of biospecimen donation and biobanking but still have a positive attitude towards biospecimen donation. Several factors, including family disease history( p <0.05), previous donation history ( p <0.01), the knowledge of biospecimen donation ( p <0.01), the knowledge of biospecimen donation conception ( p <0.01) were closely linked to donation willingness. Among those factors, family disease history, brief knowledge about biospecimen donation were independent factors affecting donation willingness. The reverse health effects and privacy leakage were major concerns among the majority of respondents. Primary reasons affected willingness, or unwillingness donation were their benefic to public interests and privacy concerns. In summary, this survey we mainly discussed factors affecting publics’ willingness and issues people concerned most in biospecimen donation of a biobank. Conclusions Most respondents hold positive attitudes towards biospecimen donation but lack relevant knowledges. Several factors influenced donation willingness probably caused by those deficiency knowledge. Even though faced this challenge, responders’ merits of altruistic behavior may contribute to the act of donation. In addition, information leakage and health impairment remained the domain factors prevented their participation. Further works are required to eliminate those undesirable elements restrained biospecimen donation by the well biobank knowledge popularization and detailed pre-donation information exchange .