Zhang Jian, Fu Yi, Lakowicz Joseph R
Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, Maryland 21201, USA.
Bioconjug Chem. 2007 May-Jun;18(3):800-5. doi: 10.1021/bc0603384. Epub 2007 Mar 22.
This work constitutes the first fluorescent imaging of cells using metal plasmon-coupled probes (PCPs) at single cell resolution. N-(2-Mercapto-propionyl)glycine-coated silver nanoparticles were synthesized by reduction of silver nitrate using sodium borohyride and then succinimidylated via ligand exchange. Alexa Fluor 647-labeled concanavalin A (con A) was chemically bound to the silver particles to make the fluorescent metal plasmon-coupled probes. The fluorescence images were collected using a scanning confocal microscopy. The fluorescence intensity was observed to enhance 7-fold when binding the labeled con A on a single silver particle. PCPs were conjugated on HEK 293 A cells. Imaging results demonstrate that cells labeled by PCPs were 20-fold brighter than those by free labeled con A.
这项工作构成了首次在单细胞分辨率下使用金属等离子体耦合探针(PCP)对细胞进行荧光成像。通过硼氢化钠还原硝酸银合成了N-(2-巯基丙酰基)甘氨酸包覆的银纳米颗粒,然后通过配体交换进行琥珀酰亚胺化。将Alexa Fluor 647标记的伴刀豆球蛋白A(Con A)化学结合到银颗粒上,制成荧光金属等离子体耦合探针。使用扫描共聚焦显微镜收集荧光图像。当在单个银颗粒上结合标记的Con A时,观察到荧光强度增强了7倍。PCP与HEK 293 A细胞共轭。成像结果表明,用PCP标记的细胞比用游离标记的Con A标记的细胞亮20倍。