Nolan John P, Duggan Erika, Condello Danilo
La Jolla Bioengineering Institute Suite 210 3535 General Atomics Court San Diego, California 92121, United States.
Bioconjug Chem. 2014 Jul 16;25(7):1233-42. doi: 10.1021/bc5000252. Epub 2014 Jun 10.
Nanoparticle surface enhanced Raman scattering (SERS) tags have attracted interest as labels for use in a variety of applications, including biomolecular assays. An obstacle to progress in this area is a lack of standardized approaches to compare the brightness of different SERS tags within and between laboratories. Here we present an approach based on binding of SERS tags to beads with known binding capacities that allows evaluation of the average intensity, the relative binding footprint of particles in a SERS tag preparation, and the size-normalized intensity or emittance. We tested this on four different SERS tag compositions and show that aggregated gold nanorods produce SERS tags that are 2-4 times brighter than relatively more monodisperse nanorods, but that the aggregated nanorods are also correspondingly larger, which may negate the intensity if steric hindrance limits the number of tags bound to a target. By contrast, SERS tags prepared from smaller gold nanorods coated with a silver shell produce SERS tags that are 2-3 times brighter, on a size-normalized basis, than the Au nanorod-based tags, resulting in labels with improved performance in SERS-based image and flow cytometry assays. SERS tags based on red-resonant Ag plates showed similarly bright signals and small footprint. This approach to evaluating SERS tag brightness is general, uses readily available reagents and instruments, and should be suitable for interlab comparisons of SERS tag brightness.
纳米颗粒表面增强拉曼散射(SERS)标签作为用于包括生物分子检测在内的各种应用的标记物已引起关注。该领域进展的一个障碍是缺乏标准化方法来比较不同实验室内部和之间不同SERS标签的亮度。在此,我们提出一种基于将SERS标签与具有已知结合能力的珠子结合的方法,该方法能够评估平均强度、SERS标签制剂中颗粒的相对结合足迹以及尺寸归一化强度或发射率。我们在四种不同的SERS标签组合物上测试了这一方法,结果表明,聚集的金纳米棒产生的SERS标签比相对更单分散的纳米棒亮2至4倍,但聚集的纳米棒相应地也更大,如果空间位阻限制了与靶标结合的标签数量,这可能会抵消强度优势。相比之下,由涂有银壳的较小金纳米棒制备的SERS标签,在尺寸归一化的基础上,比基于金纳米棒的标签亮2至3倍,从而在基于SERS的成像和流式细胞术检测中产生性能更优的标记物。基于红色共振银板的SERS标签显示出类似的明亮信号和小的足迹。这种评估SERS标签亮度的方法具有通用性,使用现成的试剂和仪器,并且应该适用于不同实验室之间SERS标签亮度的比较。