Abstract: Fluorescein methyl acrylate(FMA) was prepared using the equimolar methacryloyl chloride，triethylamine and fluorescein as raw materials. A novel kind of polymer fluorescent probes P(HPMA- FMA) were prepared using FMA and N-(2-hydroxypropyl) methacrylamide(HPMA) at the molar ratio of 1∶10 ，and 2，2′-azobis(2-methylpropio nitrile)(AIBN) as initiator． The trypan blue staining was used to assessed the cytotoxicity of P(HPMA-FMA) on HL-60 cells．Fluorescence microscopy and flow cytometry were used to make a further investigation of phagocytosis of fluorescent tracing effects by P(HPMA-FMA) on human acute promyelocytic leukemia HL-60 cells．Results indicated that the P(HPMA-FMA) was less toxic to HL-60 cells and had no effect on cell proliferation with the mass concentration of 4—16 mg/mL．The fluorescence intensity could achieve the requirement to be observed by fluorescence microscopy and detected by flow cytometry only at the level of 30 μg/mL.