Objective To investigate the value of a 4-color and 10-antibody flow cytometry immunophenotyping panel using 10 antibodies including CD45, CD38, CD19, CD56, CD20, CD5, CD10, human leukocyte antigen-DR (HLA-DR), κ antibody and λ antibody marked by four kinds of fluorescein including R-phycoerythrin (PE), fluorescein isothiocyanate (FITC), peridinin chlorophy Ⅱ protein (PerCP) and allophycocyanin (APC) in the diagnosis of multiple myeloma (MM). Methods A 4-color and 10-antibody flow cytometry immunophenotyping panel which used CD45dim/-/CD38high as gating strategy supplemented by CD19, CD56, CD20, CD10, CD5, HLA-DR, κ antibody and λ antibody was used to test the bone marrow (BM) specimens of 45 MM patients treated between December 2013 and March 2015. Then by morphological examination, we analyzed the quantitative results and characteristics of myeloma cells. Results In all the 45 MM patients, the myeloma cell detection rate was 100% by flow cytometry. The proportion range of myeloma cells in BM was between 1.17% and 72.31%, which showed a good consistency with the results of 7.5%-90.0% detected by morphological examination. The positive expression rates of antigen on myeloma cells were: 100.00% for CD38, 11.11% for CD45, 2.22% for CD19, 73.33% for CD56, 17.78% for CD20, 42.22% for HLA-DR, and 0% for CD10 and CD5. About 64.44% of the MM patients were restricted cytoplasmic λ light chain typing, and 35.56% were restricted cytoplasmic κ light chain typing. There was no obvious phenotype difference among the 3 Durie-Salmon stages of MM (P>0.05). The expression of CD56 was different among different immunoglobulin types of MM, and the types of immunoglobulin with an expression from high to low were non-secretory, IgA, IgG, and light chain (P<0.05). Conclusion The 4-color and 10-antibody flow cytometry immunophenotyping panel using 10 antibodies including CD45, CD38, CD19, CD56, CD20, CD5, CD10, HLA-DR, κ antibody and λ antibody marked by four kinds of fluorescein including PE, FITC, PerCP and APC has a good diagnostic value for MM.