Screening and diagnosing of abnormal Leukocytes are crucial for the diagnosis of immune diseases and Acute Lymphoblastic Leukemia (ALL). As the deterioration of abnormal leukocytes is mainly due to the changes in the chromatin distribution, which signi¯cantly a®ects the absorption and re°ection of light, the spectral feature is proved to be important for leukocytes classi¯cation and identi¯cation. This paper proposes an accurate identi¯cation method for healthy and abnormal leukocytes based on microscopic hyperspectral imaging (HSI) technology which combines the spectral information. The segmentation of nucleus and cytoplasm is obtained by the morphological watershed algorithm. Then, the spectral features are extracted and combined with the spatial features. Based on this, the support vector machine (SVM) is applied for classi¯cation of¯ve types of leukocytes and abnormal leukocytes. Compared with di®erent classi¯cation methods, the proposed method utilizes spectral features which highlight the differences between healthy leukocytes and abnormal leukocytes, improving the accuracy in the classi¯cation and identi¯cation of leukocytes. This paper only selects one subtype of ALL for test, and the proposed method can be applied for detection of other leukemia in the future.