Lian Wei, Litherland Sally A, Badrane Hassan, Tan Weihong, Wu Donghai, Baker Henry V, Gulig Paul A, Lim Daniel V, Jin Shouguang
Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL 32610, USA.
Anal Biochem. 2004 Nov 1;334(1):135-44. doi: 10.1016/j.ab.2004.08.005.
Fluorescent-labeled molecules have been used extensively for a wide range of applications in biological detection and diagnosis. A new form of highly luminescent and photostable nanoparticles was generated by doping the fluorescent dye tris(2'2-bipyridyl)dichlororuthenium(II)hexahydrate (Rubpy) inside silica material. Because thousands of fluorescent dye molecules are encapsulated in the silica matrix that also serves to protect Rubpy dye from photodamaging oxidation, the Rubpy-dye-doped nanoparticles are extremely bright and photostable. We have used these nanoparticles successfully in various fluorescence labeling techniques, including fluorescent-linked immunosorbent assay, immunocytochemistry, immunohistochemistry, DNA microarray, and protein microarray. By combining the high-intensity luminescent nanoparticles with the specificity of antibody-mediated recognition, ultrasensitive target detection has been achieved. In all cases, assay results clearly demonstrated the superiority of the nanoparticles over organic fluorescent dye molecules and quantum dots in probe labeling for sensitive target detection. These results demonstrate the potential to apply these newly developed fluorescent nanoparticles in various biodetection systems.
荧光标记分子已广泛应用于生物检测和诊断的众多领域。通过将荧光染料三水合二氯二（2,2'-联吡啶）钌（II）（Rubpy）掺杂到二氧化硅材料中，生成了一种新型的高发光且光稳定的纳米颗粒。由于数千个荧光染料分子被封装在二氧化硅基质中，该基质还能保护Rubpy染料免受光损伤氧化，因此掺杂Rubpy染料的纳米颗粒极其明亮且光稳定。我们已成功将这些纳米颗粒用于各种荧光标记技术，包括荧光酶联免疫吸附测定、免疫细胞化学、免疫组织化学、DNA微阵列和蛋白质微阵列。通过将高强度发光纳米颗粒与抗体介导识别的特异性相结合，实现了超灵敏的目标检测。在所有情况下，检测结果都清楚地表明，在用于敏感目标检测的探针标记方面，纳米颗粒优于有机荧光染料分子和量子点。这些结果证明了将这些新开发的荧光纳米颗粒应用于各种生物检测系统的潜力。
