Liu Yuting, Zhong Di, Yu Lei, Shi Yanfeng, Xu Yuanhong
Institute of Biomedical Engineering, College of Life Science, Basic Medical College, Qingdao University, Qingdao 266071, China.
Department of Genetics and Cell Biology, Basic Medical College, Qingdao University, No. 308 Ningxia Road, Qingdao 266000, China.
Nanomaterials (Basel). 2023 Jan 21;13(3):437. doi: 10.3390/nano13030437.
Small molecular dyes are commonly used for bacterial imaging, but they still meet a bottleneck of biological toxicity and fluorescence photobleaching. Carbon dots have shown high potential for bio-imaging due to their low cost and negligible toxicity and anti-photobleaching. However, there is still large space to enhance the quantum yield of the carbon quantum dots and to clarify their mechanisms of bacterial imaging. Using carbon dots for dyeing alive bacteria is difficult because of the thick density and complicated structure of bacterial cell walls. In this work, both dead or alive bacterial cell imaging can be achieved using the primary amine functionalized carbon dots based on their small size, excellent quantum yield and primary amine functional groups. Four types of carbon quantum dots were prepared and estimated for the bacterial imaging. It was found that the spermine as one of precursors can obviously enhance the quantum yield of carbon dots, which showed a high quantum yield of 66.46% and high fluorescence bleaching-resistance (70% can be maintained upon 3-h-irradiation). Furthermore, a mild modifying method was employed to bound ethylenediamine on the surface of the spermine-carbon dots, which is favorable for staining not only the dead bacterial cells but also the alive ones. Investigations of physical structure and chemical groups indicated the existence of primary amine groups on the surface of spermine-carbon quantum dots (which own a much higher quantum yield) which can stain alive bacterial cells visibly. The imaging mechanism was studied in detail, which provides a preliminary reference for exploring efficient and environment-friendly carbon dots for bacterial imaging.
小分子染料常用于细菌成像,但它们仍面临生物毒性和荧光光漂白的瓶颈。碳点由于其低成本、可忽略的毒性和抗光漂白性,在生物成像方面显示出巨大潜力。然而,提高碳量子点的量子产率并阐明其细菌成像机制仍有很大空间。由于细菌细胞壁密度大、结构复杂,使用碳点对活细菌进行染色很困难。在这项工作中,基于其小尺寸、优异的量子产率和伯胺官能团,使用伯胺功能化的碳点可以实现死细菌或活细菌细胞成像。制备了四种类型的碳量子点并对其进行细菌成像评估。发现精胺作为前体之一可以显著提高碳点的量子产率,其量子产率高达66.46%,并且具有高抗荧光漂白性(在3小时照射后可保持70%)。此外,采用了一种温和的修饰方法将乙二胺结合到精胺碳点表面,这不仅有利于对死细菌细胞染色,也有利于对活细菌细胞染色。对物理结构和化学基团的研究表明,精胺碳量子点(其量子产率高得多)表面存在伯胺基团,该基团可以明显地对活细菌细胞进行染色。对成像机制进行了详细研究,为探索用于细菌成像的高效且环境友好的碳点提供了初步参考。