微波合成靶向核仁的荧光碳纳米颗粒研究  

Microwave-synthesized Fluorescent Carbon Nanoparticles for Nucleolus Imaging

作　　者：王倩[1] 冯真真 毛健 刘立华 陈晓良 魏晓贇 黄方[1] WANG Qian;FENG Zhen-Zhen;MAO Jian;LIU Li-Hua;CHEN Xiao-Liang;WEI Xiao-Yun;HUANG Fang(State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology,China University of Petroleum(East China),Qingdao 266580,China;Testing Institute for Chemicals&Minerals,Shandong Entry-Exit Inspection and Quarantine Bureau,Qingdao 266500,China)

机构地区：[1]中国石油大学(华东)重质油国家重点实验室和生物工程与技术中心,青岛266580 [2]山东省出入境检验检疫局化工矿物测试所,青岛266500

出　　处：《生物化学与生物物理进展》2021年第10期1204-1213,共10页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金(05E09040310,21874154,42061134020);中央高校基础科研业务费专项基金(18CX02126A);青岛海关研究项目(QK202035)资助。

摘　　要：核仁是细胞内重要的亚核结构,其在恶性病的演变过程中扮演重要角色,是病理学家诊断癌症的重要指标.尽管核仁如此关键,但到目前为止,核仁的荧光探针寥寥无几.本文以水杨酸和1,8-二氨基萘为反应物,通过微波消解法合成了一种新型荧光碳纳米颗粒(FCNs),采用透射电子显微镜、动态光散射仪、傅里叶红外光谱仪、紫外分光光度计、荧光光谱仪等对其物理、化学、光学性质进行了表征、分析.借助激光扫描共聚焦等技术对FCNs的细胞摄取机制及分布进行了探究.实验结果表明,所合成碳纳米颗粒尺寸均匀,最佳激发波长在348 nm,对应的最大发射峰为432 nm,荧光量子产率为17.8%,荧光寿命为1.13 ns,其表面含有丰富的氨基和羟基,光稳定性强且毒性极低,可实现对细胞核仁染色,并且随着共孵育时间的延长,进入细胞的量越多,靶向核仁更明显.此外,经过对FCNs细胞摄取路径的考察,发现FCNs是通过小窝介导的路径被内吞.该研究为碳基纳米材料在亚细胞器靶向成像的应用方面提供了有力的工具和新思路.The nucleolus is an important subnuclear structure in the cell,which plays an important role in the evolution of malignancies and the diagnosis of cancer.Although the nucleolus is critical,so far,there are really few fluorescent probes for the nucleolus.In this paper,salicylic acid and 1,8-diaminonaphthalene were used as carbon and nitrogen sources,and a novel fluorescent carbon nanoparticles(FCNs)were synthesized by microwave digestion system.Its physical,chemical and optical properties of FCNs were characterized and analyzed by using transmission electron microscope(TEM),dynamic light scattering instrument(DLS),Fourier infrared spectrometer,ultraviolet spectrophotometer,fluorescence spectrometer,etc.The experimental results demonstrated that the synthesized carbon nanoparticles were uniform in size and rich with amino groups on surface.Its best excitation wavelength was 348 nm,the corresponding maximum emission peak was 432 nm.Its fluorescence quantum yield was 17.8%and fluorescence lifetime was 1.13 ns.Moreover,the photostability and cytotoxicity of FCNs were investigated with illumination and MTT assays.Our results suggest that FCNs exhibit to be a really photostable and low toxic probe.To confirm the potential application of this fluorescence FCNs in bioimaging,the FCNs were designed to stain human cervical cancer He La cells.After incubation,we found that FCNs enable the function of selectively staining the nucleolus of living cells due to its positive chargeability.Its cellular uptake mechanism and intracellular distribution in He La cells were explored further by using confocal laser scanning microscopy and TEM.It was interesting to observe that more FCNs entered the cell,and lighted the nucleolus more obviously with the extension of the co-incubation time.In addition,after investigating the cells’uptake path of FCNs,we found that FCNs were endocytosed via caveolae-mediated pathway rather than clathrin mediated pathways or macropinocytosis.Our study provided a potential probe for nucleolus and this find

关 键 词：碳基纳米材料 荧光性能 核仁靶向 

分 类 号：Q6[生物学—生物物理学]