Zhang Jian, Fu Yi, Liang Dong, Zhao Richard Y, Lakowicz Joseph R
Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, Maryland 21201, USA.
Langmuir. 2008 Nov 4;24(21):12452-7. doi: 10.1021/la801749f. Epub 2008 Oct 7.
Silver island films (SIFs) were deposited on glass substrates to serve as supports. T-Lymphocytic (PM1) cell lines were labeled by Alexa Fluor 680-dextran conjugates on the membranes or by YOYO in the nuclei. The fluorescence images of the cell lines were recorded in the emission intensity and lifetime using scanning confocal microscopy. The fluorescence signals by the fluorophores bound on the cell membranes were enhanced significantly by SIF supports as compared with those on the glass. In addition to the increase in the intensity, there was a dramatic shortening of the emission lifetime. In contrast to the Alexa Fluor 680 fluorophores on the membranes, the YOYO fluorophores intercalated in the cell nuclei were not influenced significantly by the silver islands. This result can be interpreted by an effect of the distance on coupling between the fluorophores and metal particles: the fluorophores on the cell membranes are localized within, but the fluorophores in the cell nuclei are beyond the region of metal-enhanced fluorescence. Thus, the metal supports can be used to improve the detection sensitivity for target molecules on cell surfaces when they are fluorescently labeled.
银岛膜(SIFs)沉积在玻璃基板上作为支撑物。T淋巴细胞(PM1)细胞系在细胞膜上用Alexa Fluor 680 - 葡聚糖共轭物标记,在细胞核中用YOYO标记。使用扫描共聚焦显微镜记录细胞系的荧光图像的发射强度和寿命。与玻璃上的情况相比,SIF支撑物显著增强了结合在细胞膜上的荧光团的荧光信号。除了强度增加外,发射寿命也显著缩短。与细胞膜上的Alexa Fluor 680荧光团不同,插入细胞核中的YOYO荧光团受银岛的影响不显著。这一结果可以通过荧光团与金属颗粒之间耦合的距离效应来解释:细胞膜上的荧光团位于金属增强荧光区域内,但细胞核中的荧光团在该区域之外。因此,当目标分子用荧光标记时,金属支撑物可用于提高细胞表面目标分子的检测灵敏度。