Objective To evaluate the diagnostic value of the cell block （CB） with immunostaining method by endoscopic ultrasound-guided fine-needle aspiration （EUS-FNA） biopsy for pancreatic lesions. Methods A total of 72 patients with pancreatic lesions underwent EUS-FNA at the First Affiliated Hospital of Guangxi Medical University from March 2012 to June 2013. The EUS-FNA samples of all patients were processed by conventional smear cytology, liquid-based cytology （LBC） and the cell block with immunos- taining. Results There were 61 pancreatic patients who were finally diagnosed as having pancreatic tumors, including 55 cases of pancreatic cancer, 2 pancreatic solid pseudopapillary tumor, 4 pancreatic endocrine tumors （PETs） , and 11 benign lesions： 4 chronic pancreatitis, 2 pancreatic tuberculosis ,4 pancreatitis and 1 pancreatic mucinous cystadenoma. The diagnostic sensitivity of conventional smear cytology, liquid-based cytology and cell block with immuno-staining method were 68. 9% （42/61）, 75.4% （46/61） and 90. 2% （ 55/61 ） , respectively. The diagnostic specificity of three methods were all 100.0%. The diagnostic accura- cy rates were 73.6% （53/72） ,79. 2% （57/72） and 91.7% （66/72）, respectively. The diagnostic accuracy rate of the cell block with immunostaining was higher than those of conventional smear cytology （ P 〈 0. 05） and the liquid-based cytology （P 〈 0. 05 ）. Conclusion Endoscopic ultrasound-guided fine-needle aspiration is a safe and effective method with high sensitivity and specificity in the diagnosis of pancreatic lesions. Cell block method combining immunohistochemistry helps to increase the diagnosis and histological diagnosis of pancreatic lesions. The cell block has a greater clinical value in the diagnosis of pancreatic lesions.