State Key Laboratory for Structural Chemistry of Unstable and Stable Species, Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, 100190 Beijing, China; Graduate School, University of Chinese Academy of Sciences, 100049 Beijing, China.
State Key Laboratory for Structural Chemistry of Unstable and Stable Species, Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, 100190 Beijing, China.
Biosens Bioelectron. 2014 May 15;55:76-82. doi: 10.1016/j.bios.2013.12.003. Epub 2013 Dec 10.
Due to mitochondria involved in both apoptotic and necrotic cell death, labeling and imaging mitochondria has attracted considerable interest. However, conventional organic dyes used for mitochondrial imaging are limited because of their poor photostability. Considering that gold nanoclusters (AuNCs) possess some advantages over considerable interest, such as excellent photostability and strong fluorescence emission, we herein prepared a mitochondria-targeted fluorescent probe, AuNCs@CS-TPP, based on a covalent link between triphenylphosphonium (TPP) cations and chitosan-coated AuNCs (AuNCs@CS). The as-prepared AuNCs@CS-TPP exhibited a bluish fluorescence emission at 440 nm with a quantum yield of 8.5%. Meanwhile, the fluorescence intensity of AuNCs@CS-TPP labeled HeLa cells did not show apparent decrease after 8 min irradiation. Cytotoxicity assay showed that AuNCs@CS-TPP did not display any appreciable cytotoxicity on cells even at a concentration of 60 μg mL(-1). In addition, the result of fluorescence co-localization imaging in vitro indicated that AuNCs@CS-TPP could selectively accumulate into mitochondria of HeLa cells and HepG2 cells. These findings demonstrated that AuNCs@CS-TPP possessed superior photostability, low cytotoxicity, high sensitivity and target-specificity to mitochondria, allowing labeling and imaging of the mitochondria in living cells.
由于线粒体参与细胞凋亡和坏死,因此标记和成像线粒体引起了相当大的兴趣。然而,传统的用于线粒体成像的有机染料由于其较差的光稳定性而受到限制。考虑到金纳米簇(AuNCs)具有一些优势,例如出色的光稳定性和强荧光发射,我们在此基于三苯基膦(TPP)阳离子与壳聚糖包覆的 AuNCs(AuNCs@CS)之间的共价键合,制备了一种靶向线粒体的荧光探针 AuNCs@CS-TPP。所制备的 AuNCs@CS-TPP 在 440nm 处表现出蓝色荧光发射,量子产率为 8.5%。同时,AuNCs@CS-TPP 标记的 HeLa 细胞在 8 分钟照射后荧光强度没有明显下降。细胞毒性测定表明,即使在 60μgmL(-1)的浓度下,AuNCs@CS-TPP 对细胞也没有显示出任何明显的细胞毒性。此外,体外荧光共定位成像的结果表明,AuNCs@CS-TPP 可以选择性地积累到 HeLa 细胞和 HepG2 细胞的线粒体中。这些发现表明,AuNCs@CS-TPP 具有优越的光稳定性、低细胞毒性、对线粒体的高灵敏度和靶向特异性,允许对活细胞中的线粒体进行标记和成像。