Neutrophil surface molecules function in part as biological sensors. Surface antigens undergo several changes during myeloid differentiation to accommodate cell functions. Here we describe time-dependent changes in antigen expression at the surface of HL-60 myeloid cells when differentiation is induced into granulocytes by dimethylsulphoxide (DMSO), all-trans retinoic acid (ATRA), as well as PLB-985 myeloid cells induced to differentiate into neutrophils by a mixture of N,N-Dimethylformamide (DMF) and Nutridoma-SP. Terminally differentiated cells were compared to circulating neutrophils in terms of cell surface antigen expression by flow cytometry analysis using a panel of pertinently purported 35 antibodies. The changes in myeloid cell surface antigens induced by DMSO, ATRA or DMF/Nutridoma-SP paralleled the expression pattern of these molecules in normal granulopoeisis with the exception of 7 antigens up-regulated and 9 down-regulated, indicating that the maturation was most probably not achieved and thus partially defective. All these differentiation inducers failed to induce the expression of neutrophil specific markers CD16 and CD66b. Differentiated PLB-985 cells appeared closer to neutrophils, in term of maturation and CD expression than the DMSO- or ATRA-differentiated HL-60. Finally, single cell analysis of the expression dynamics of several differentiation markers (CD11b, CD14, CD35, CD71) revealed a bistable switch and not a graded change of expression when measured as a cell population average. These results demonstrate kinetic changes in cell surface antigen expression during the transformation of a proliferating leukaemic cell into a potentially mature and terminally differentiated cell, which might be of substantial importance for analyzing the gene expression pathways that govern granulopoiesis.