Abstract： Objective To evaluate the utility of flow cytometry (FCM) in diagnosis and subclassification of non-Hodgkin lymphoma (NHL). Methods The samples of lymph nodes biopsy from 59 cases clinically suspected of NHL were detected by flow cytometry; and clonal lymphocytes and their immunophenotypes were identified analyzed. The concordance between the results of flow cytometry and histopathology was analyzed. Results Among the 59 cases, flow cytometry was able to identify aberrant clonal lymphocytes in 24 of 28 NHL cases identified by histopathology, the neoplastic lymphocytes ranged from 4.28 % to 89.10 %; 23 cases were diagnosed as B-NHL and 1 case was diagnosed as T-NHL. Compared with histopathology, the accuracy of FCM was 85.71% in diagnosis of NHL. The specificity and sensitivity of FCM was 100 % and 92% in diagnosis of B-NHL. The accuracy of flow cytometry immunophenotyping in classification of 24 cases of NHL was consistent with that of histopathology. Conclusion Flow cytometry could be an ancillary technique in diagnosis of NHL by identifying aberrant clonal lymphocytes, and enable identification of B-NHL subtype.