Jaworska Aleksandra, Wojcik Tomasz, Malek Kamilla, Kwolek Urszula, Kepczynski Mariusz, Ansary Abu A, Chlopicki Stefan, Baranska Malgorzata
Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Bobrzynskiego 14, 30-348 Krakow, Poland ; Faculty of Chemistry, Jagiellonian University, Ingardena 3, 30-060 Krakow, Poland.
Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Bobrzynskiego 14, 30-348 Krakow, Poland.
Mikrochim Acta. 2015;182(1):119-127. doi: 10.1007/s00604-014-1307-5. Epub 2014 Jun 19.
Fluorescence and surface-enhanced Raman scattering (SERS) spectroscopy were employed to investigate the cellular uptake of rhodamine 6G (R6G) alone and of R6G loaded with gold nanoparticles (AuNPs) by endothelial cells. R6G plays the role of a Raman reporter in SERS but also displays strong fluorescence. The presence of bare R6G molecules and R6G-AuNPs in the cytoplasm of the cells is detected via the 2D fluorescence of the dye after a 0.5 h of the incubation with R6G and R6G-AuNPs, and then the concentration of the dye increases within 4 h of exposure. The examination of the cellular uptake of the R6G and R6G-AuNPs species at different temperatures suggests that the internalization of the R6G-AuNPs into endothelial cells occurs mainly via endocytosis. 3D fluorescence imaging of R6G inside cells reveals inhomogeneous distribution of the dye in the cytoplasm. The SERS signal of the Raman reporter inside the cell disappears after 2 h of incubation with R6G-AuNPs and then amino acid residues, purines and pyrimidines become SERS-active via their interactions with the gold. The results highlight the significance of using multiple techniques to cover a spectrum of issues in the application of SERS nanosensors for probing an intracellular environment under comparable and standardized conditions. FigureCellular uptake of bare rhodamine 6G and rhodamine 6G adsorbed onto AuNPs were studied on endothelial cells using fluorescence and surface-enhanced Raman spectroscopy. The internalization of R6G-AuNPs occurs via endocytosis and diffusion resulting in uneven distribution in the cytoplasm.
采用荧光和表面增强拉曼散射（SERS）光谱法研究了内皮细胞对单独的罗丹明6G（R6G）以及负载金纳米颗粒（AuNPs）的R6G的细胞摄取情况。R6G在SERS中起拉曼报告分子的作用，但也显示出强烈的荧光。在用R6G和R6G-AuNPs孵育0.5小时后，通过染料的二维荧光检测细胞胞质中裸露的R6G分子和R6G-AuNPs的存在，然后在暴露4小时内染料浓度增加。在不同温度下对R6G和R6G-AuNPs种类的细胞摄取检查表明，R6G-AuNPs内化进入内皮细胞主要通过内吞作用。细胞内R6G的三维荧光成像揭示了染料在细胞质中的分布不均匀。在用R6G-AuNPs孵育2小时后，细胞内拉曼报告分子的SERS信号消失，然后氨基酸残基、嘌呤和嘧啶通过与金的相互作用变得具有SERS活性。结果突出了在可比和标准化条件下使用多种技术来涵盖SERS纳米传感器在探测细胞内环境应用中的一系列问题的重要性。图 使用荧光和表面增强拉曼光谱研究了内皮细胞对裸露的罗丹明6G和吸附在AuNPs上的罗丹明6G的细胞摄取。R6G-AuNPs的内化通过内吞作用和扩散发生，导致在细胞质中分布不均匀。
