Single cell analysis using surface enhanced Raman scattering (SERS) tags.
Single cell analysis using surface enhanced Raman scattering (SERS) tags.
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DOI：
10.1016/j.ymeth.2012.03.024
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发表时间：
2012-07
期刊：
影响因子：
4.8
通讯作者：
Stoner, Samuel A.
中科院分区：
文献类型：
--
作者：
Nolan, John P.;Duggan, Erika;Liu, Er;Condello, Danilo;Dave, Isha;Stoner, Samuel A.
荧光是生物分析方法的支柱，提供敏感和定量报告，通常用于多路或多参数分析。也许后者的最佳例子是流式细胞术，其中配备了多个激光器和检测器的仪器可以同时测量15个或更多不同的荧光团，但超过这个数字的增加受到相对较宽的发射光谱的限制。金属纳米颗粒的表面增强拉曼散射（SERS）可以产生与荧光相媲美的信号强度，但具有更窄的光谱特征，允许更大程度的复用。我们正在开发纳米粒子SERS标签以及拉曼流式细胞仪，用于悬液或贴壁细胞的多参数单细胞分析。SERS标签是基于等离子体活性纳米粒子（金纳米棒），其等离子体共振可以在所需的激发波长下调谐以提供最佳的SERS信号。拉曼共振化合物被吸附在纳米颗粒上，赋予每个SERS标签独特的光谱指纹，然后将其封装在聚合物涂层中，用于与抗体或其他靶向分子结合。拉曼流式细胞术采用高分辨率光谱流式细胞仪，能够测量完整的SERS光谱，以及传统的流式细胞术测量，每分钟数千个单个细胞。自动光谱分解算法从每个细胞中提取每个SERS标签的贡献，生成高含量、多参数的单细胞种群数据。基于sers的细胞术是对传统荧光细胞术的有力补充。SERS信号的窄光谱特征使得使用单个激光器和探测器在更小的光谱区域内测量更多不同的探头，从而允许更高水平的多路复用和多参数分析。
Fluorescence is a mainstay of bioanalytical methods, offering sensitive and quantitative reporting, often in multiplexed or multiparameter assays. Perhaps the best example of the latter is flow cytometry, where instruments equipped with multiple lasers and detectors allow measurement of 15 or more different fluorophores simultaneously, but increases beyond this number are limited by the relatively broad emission spectra. Surface enhanced Raman scattering (SERS) from metal nanoparticles can produce signal intensities that rival fluorescence, but with narrower spectral features that allow a greater degree of multiplexing. We are developing nanoparticle SERS tags as well as Raman flow cytometers for multiparameter single cell analysis of suspension or adherent cells. SERS tags are based on plasmonically active nanoparticles (gold nanorods) whose plasmon resonance can be tuned to give optimal SERS signals at a desired excitation wavelength. Raman resonant compounds are adsorbed on the nanoparticles to confer a unique spectral fingerprint on each SERS tag, which are then encapsulated in a polymer coating for conjugation to antibodies or other targeting molecules. Raman flow cytometry employs a high resolution spectral flow cytometer capable of measuring the complete SERS spectra, as well as conventional flow cytometry measurements, from thousands of individual cells per minute. Automated spectral unmixing algorithms extract the contributions of each SERS tag from each cell to generate high content, multiparameter single cell population data. SERS-based cytometry is a powerful complement to conventional fluorescence-based cytometry. The narrow spectral features of the SERS signal enables more distinct probes to be measured in a smaller region of the optical spectrum with a single laser and detector, allowing for higher levels of multiplexing and multiparameter analysis.
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