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二苯并类双光子材料的制备及作为生物探针的基础研究
其他题名Preparation of dibenzo-based two-photon materials and their basic research as biological probes
迟书恒
学位类型博士
导师吴谊群
2017
学位授予单位中国科学院上海光学精密机械研究所
关键词双光子荧光探针 光开关效应 细胞核定位 光稳定性
摘要本文以芴酮、二苯并噻吩和二苯并呋喃为中心共轭桥制备了10种未见报道的吡啶阳离子衍生物,研究了新化合物在N,N-二甲基甲酰胺(DMF)中的单光子光学性质,包括线性吸收光谱、荧光光谱和荧光量子产率。采用脉宽为120 fs、波长为800 nm和重复频率为1 kHz(Ti:蓝宝石激光器)的脉冲激光研究了新化合物的双光子光学性质,采用参比法计算了不同波长下新化合物在DMF溶液中的双光子吸收截面;研究了目标化合物和小牛胸腺DNA在Tris-HCl缓冲溶液中相互作用前后的吸收光谱、单光子荧光光谱和双光子荧光光谱变化。讨论了化学结构对新材料单、双光子光学性质和光开关效应的影响。 以新材料作为生物荧光探针染色小鼠成肌细胞3T3,利用德国Carl Zeiss LSM710激光共聚焦荧光显微镜对染色样品进行了单、双光子荧光成像研究。通过新材料与商用细胞核染色剂Hochest 33342的共染色成像实验,研究了新材料作为荧光探针的核定位能力;通过新材料与商用线粒体染色剂MTR的双染实验,研究了新材料在细胞多色标记方面的膜渗透性和复染相容性;研究了不同渗透深度下新材料与商用核定位剂DAPI在3T3细胞中的三维成像能力。在波长分别为800 nm和740 nm的双光子共聚焦激光的持续辐照下,研究了新材料和DAPI在3T3细胞中的双光子荧光强度随时间的变化趋势;比较和分析了从荧光发射到熄灭过程中新材料和DAPI的荧光持续时间、荧光衰减速率和荧光衰减幅度,讨论了化学结构对新材料光稳定性的影响。
其他摘要In this paper, a series of novel pyridine cationic deriatives containing fluorenone, dibenzothiophene and dibenzofuran as the central π-conjuated bridges were reported. The one-photon optical properties of new compounds in N,N-dimethylfomamide solution were studied, including linear absorption spectra, fluorescence spectra and fluorescence quantum yields. The two-photon optical properties of new compounds were studied by pulse laser (Ti: sapphire) with a pulse width of 120 fs, a wavelength of 800 nm and a repetition rate of 1 kHz. The two-photon absorption cross-sections of new compounds in DMF solution at different wavelengths were calculated by using the reference method; The absorption spectra, one-photon fluorescence spectra and two-photon fluorescence spectra of the target compounds and calf thymus DNA before and after the interaction in Tris-HCl buffer solution were studied. The influence of chemical structure on the one and two-photon optical properties and fluorescence “off-on” effects of new materials were discussed. The new materials were used as biological fluorescent probes to stain the 3T3-Swiss albino cells, and the one and two-photon fluorescence imaging experiments were performed with the Zeiss LSM710 Carl laser confocal fluorescence microscope. The nuclear localization ability of the new materials were studied by the co-staining experiment between the new material and the commercial nuclear staining reagent Hochest 33342; The membrane permeability and the counterstain compatibility of the new materials in multicolor cell labeling application were studied by means of the double-staining experiments between the new materials and the commercial mitochondrial dye MTR; Three dimensional imaging capabilities of new materials and commercial nuclear staining agent DAPI in 3T3 cells under different penetration depths were studied. Under continuous irradiation at wavelengths of 800 nm and 740 nm, respectively, the change trends of the time-dependent two-photon fluorescence intensity of new materials and DAPI in 3T3 cells were studied; the fluorescence duration, fluorescence decay rate and fluorescence decay amplitude of new materials and DAPI were compared and analyzed, and the influences of chemical structure on the photostability of new materials were also discussed.
语种中文
文献类型学位论文
条目标识符http://ir.siom.ac.cn/handle/181231/16001
专题学位论文
推荐引用方式
GB/T 7714
迟书恒. 二苯并类双光子材料的制备及作为生物探针的基础研究[D]. 中国科学院上海光学精密机械研究所,2017.
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