Progress in the management and understanding of acute leukaemia can only be obtained if these diseases are thoroughly investigated, both clinically and with a series of biological tools. This alone has made and still will make possible the identification of prognostic factors and of useful markers for the follow-up of patients in remission. Among the variety of approaches of acute leukaemia definition, immunophenotyping has taken over the past 25 years a predominant and now well-defined place, although room is left for further improvement. In this review, the current state-of-the-art of immunophenotyping of acute leukaemias will be replaced in the context of physiological leukocyte maturation. The recognized classifications and recommended immunophenotyping panels will then be discussed, and the clinical relevance of several key features will be presented. Finally, more recent openings for the use of immunophenotyping will be evoked.