Lin Juqiang, Chen Rong, Feng Shangyuan, Li Yongzeng, Huang Zufang, Xie Shusen, Yu Yun, Cheng Min, Zeng Haishan
Key Laboratory of Optoelectronic Science and Technology for Medicine, Ministry of Education and Fujian Provincial Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou 350007, China.
Biosens Bioelectron. 2009 Oct 15;25(2):388-94. doi: 10.1016/j.bios.2009.07.027. Epub 2009 Aug 3.
In current intracellular surface-enhanced Raman spectroscopy (SERS) measurements, gold or silver nanoparticles are delivered into living cells by "passive uptake". This procedure is time-consuming, could take up to several to twenties hours of incubation with nanoparticles in the culture medium. It is a less optimal method for certain applications such as high-throughput disease screening. Here, we present a method based on electroporation for fast delivery of silver nanoparticles into living cells for intracellular SERS spectroscopy. This new method for nanoparticle delivery averts the shortcoming of "passive uptake" and allows for quick acquisition of robust SERS spectra from living C666, A431, and CA46 cancer cell lines in our study. Our study also shows that the silver nanoparticles are localized only in the cell cytoplasm for electroporation delivery, while for "passive uptake", the nanoparticles have gone beyond the cytoplasm and into the nucleus. However, the whole-cell detection SERS spectra using electroporation delivery are more reproducible than for "passive uptake", thus are favored for practical applications. As a result, the process of SERS detection is accelerated significantly and the data reproducibility is improved as well, demonstrating great potential for biomedical applications, such as for high-throughput cancer cell screening.
在当前的细胞内表面增强拉曼光谱（SERS）测量中，金或银纳米颗粒通过“被动摄取”被递送至活细胞中。此过程耗时较长，在培养基中与纳米颗粒孵育可能需要数小时至二十小时。对于某些应用，如高通量疾病筛查而言，这并非最佳方法。在此，我们提出一种基于电穿孔的方法，用于将银纳米颗粒快速递送至活细胞中以进行细胞内SERS光谱分析。这种纳米颗粒递送新方法避免了“被动摄取”的缺点，并使我们能够快速从活的C666、A431和CA46癌细胞系中获取可靠的SERS光谱。我们的研究还表明，对于电穿孔递送，银纳米颗粒仅定位于细胞质中，而对于“被动摄取”，纳米颗粒已超出细胞质进入细胞核。然而，使用电穿孔递送的全细胞检测SERS光谱比“被动摄取”更具可重复性，因此更适合实际应用。结果，SERS检测过程显著加速，数据可重复性也得到提高，显示出在生物医学应用中的巨大潜力，如高通量癌细胞筛查。
