Acute leukemias of ambiguous lineage are a heterogenous group of diseases that include acute undifferentiated leukemias and mixed-phenotype acute leukemias (MPALs). These leukemias pose a challenge for pathologists and clinicians alike in diagnosis, treatment, and further management. Recent genetic characterization has provided insights into their underlying biology and classification, and has offered potential for targeted therapies. This article addresses diagnosis of MPALs with examples of the most common pitfalls, recent comprehensive molecular studies, and advancement in treatment and follow-up modalities.