Single-target molecule detection with nonbleaching multicolor optical immunolabels
Single-target molecule detection with nonbleaching multicolor optical immunolabels
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DOI：
10.1073/pnas.97.3.996
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发表时间：
2000-02-01
影响因子：
11.1
通讯作者：
Schultz, DA
中科院分区：
文献类型：
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作者：
Schultz, S;Smith, DR;Schultz, DA
我们介绍和演示了胶体银等离子体共振粒子(PRPS)在典型的生物检测中作为光学记录器的使用。PRPS是一种超亮、纳米尺寸的光学散射体，它弹性地散射光，并可以在可见光光谱中的任何颜色处制备散射峰。利用配置为暗场显微镜的显微镜，以及典型功率的白光照明，可以很容易地单独观察PRP。在这里，我们说明了使用表面涂有标准配体的PRPS作为原位杂交和免疫细胞学分析中的靶标。我们认为，PRPS可以取代或补充已建立的标记，如基于放射性、荧光、化学发光或酶/比色检测的标记，这些标记通常用于生物化学、细胞生物学和医学诊断应用。此外，由于PRP标记不会漂白，而且足够明亮，可以快速识别和计数，因此基于单目标分子检测的超灵敏分析形式现在是可行的。我们还给出了山羊抗生物素抗体的模型夹心免疫分析的结果，其中图像中计数的PRP标记的数量构成了测量信号。
We introduce and demonstrate the use of colloidal silver plasmon-resonant particles (PRPs) as optical reporters in typical biological assays. PRPs are ultrabright, nanosized optical scatterers, which scatter light elastically and can be prepared with a scattering peak at any color in the visible spectrum. PRPs are readily observed individually with a microscope configured for dark-field microscopy, with white-light illumination of typical power. Here we illustrate the use of PRPs, surface coated with standard ligands, as target-specific labels in an in situ hybridization and an immunocytology assay. We propose that PRPs can replace or complement established labels, such as those based on radioactivity, fluorescence, chemiluminescence, or enzymatic/colorimetric detection that are used routinely in biochemistry, cell biology, and medical diagnostic applications. Moreover, because PRP labels are non-bleaching and bright enough to be rapidly identified and counted, an ultrasensitive assay format based on single-target molecule detection is now practical. We also present the results of a model sandwich immunoassay for goat anti-biotin antibody, in which the number of PRP labels counted in an image constitutes the measured signal.