As one of efficient analytes, fluorescent microspheres have shown much usability on many biochemical and biomedical processes. Recent applications with fluorescent microspheres have included cytokine quantitation, single nucleotide polymorphism genotyping, phosphorylated protein detection, and characterization of the molecular interaction of nuclear receptors. These,coupled with the rapid advances in molecular biology and synthesis techniques of drugs, have presented a basis for drug screening in a high-throughput format. Based on fluorescent microspheres,earlier assay formats of HTS relied mainly on proximity-dependent energy transfer including scintillation proximity assay (SPA) (Amersham Pharmacia Biotech) and FlashPlatesTM (NEN Life Science Products, Boston, MA). Indeed, drug screening-based such fluorescent emission is still accounting for about 20～50% of current content of high-throughput screening (HTS). Now, SPA is almost a standard technique in common HTS-lab. In literature, SPA microspheres is generally prepared from inorganic scintillators such as yttrium silicate and hydrophobic polymers such as polyvinyl toluene. However, in HTS research, such microspheres often show the disadvantages of strong hydrophobicity and low quantum efficiency. The strong hydrophobicity is mainly attributed to the hydrophobic monomer, vinyl toluene. The low quantum efficiency can be as a result of low transparence of the polymer, polyvinyl toluene. Thus, the subsequent treatments for such microspheres, so as coat a polyhydroxy film to decrease the hydrophobicity, are actually considerably complicated.It has been well known that poly(methyl methacrylate) (PMMA), a good biocompatible polymer with not only adequate mechanical strength but also excellent transparence, can be regarded as an ideal candidate material for fluorescent matrix. In present study, methyl methacrylate as monomer and 2,5-diphenyloxazole (DPO) as fluorescent dye were used to the fluorescent microspheres. In guaranteeing the hydrophilicity of microsphere surfaces, dispersion polymerization was in common use (stabilizer, Polyvinyl pyrrolidone). As is apparent, with such a method in hand, one would normally find that almost no subsequent treatment for microspheres can be involved. Also such a novel fluorescent microspheres is a more suitable and the method used in present research is more practical method in comparison to common method. As the original work, the purposes of this article are to synthesize such fluorescent microspheres and probe the probable roles of synthesis conditions on microsphere synthesis. The effects of stabilizers, initiators, dispersion mediums, monomer content, and reaction temperature on the synthesis process and the particle size, as well as its panicle distribution have been shown. Also the probable role of fluorescent dye in the polymerization has been thermodynamicaily discussed.
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