Leukemia, the most common childhood cancer, is predominantly acute, with acute lymphoblastic leukemia (ALL) and acute myelogenous leukemia (AML) being the primary subtypes. The incidence rates for ALL and AML are 1.5 and 2.7 per 100,000 population, respectively. The objective of this study conducted at the Pathology department of M.G.M. Medical College in Indore was two-fold: firstly, to classify acute leukemia into ALL or AML using flow cytometry, and secondly, to assess the diagnostic value of specific markers (Anti-MPO, CD34, HLA-DR, CD45, CD79a, CD3) in acute leukemia diagnosis. The study involved evaluating cases through routine peripheral smear and subsequently analyzing relevant cases on a flow cytometer. The results revealed that 23 cases (77%) were diagnosed as ALL, while 7 cases (23%) were identified as AML. Among the ALL cases, 19 were classified as B-cell ALL (64%), whereas 4 were T-cell ALL (13%). In conclusion, the use of multicolor flow cytometry enables simultaneous assessment of multiple antigens, facilitating more precise categorization of cell populations. Furthermore, flow cytometry plays a crucial role in differentiating AML from ALL and its subtypes.