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Nucleoside-Based Ultrasensitive Fluorescent Probe for the Dual Mode Imaging of Microviscosity in Living Cells

机译:基于核苷的超灵敏荧光探针在活细胞中微粘度的双模式成像

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Microviscosity changes of living cells have a far-reaching influence on diffusion and movement capacity of RNA and, more seriously, could modify RNA functions in living cells. Fluorescent rotor, whose fluorescence responds to different environmental viscosities, holds great potential for the imaging of viscosity in biosystem. Although many fluorescent rotors have been reported for viscosity, the fluorogenic rotor with ultrasensitivity for the determination of microviscosity (<10 cP) was rarely reported. Herein, we report a nucleoside-based two-photon fluorescent rotor (dABp-3) that can selectively and ultrasensitively image microviscosity in RNA region of living cells for the first time. 2'-Deoxyadenosine is selected as an electron donor to permit energy transfer via the acetylenic bond to acceptor, a typical boron dipyrromethene moiety. Another highlight, dABp-3 is based on 2'-deoxyadenosine, which result in its recognition capacity for RNA. dABp-3 with ultrasensitivity provides a varied linear response to the microrange viscosity (1.8-6.0 cP) in RNA region of living cells on dual-mode two-photon ratio mode and fluorescence lifetime mode. After screening and optimization, advantageously, dABp-3 can be used to screen reticulocytes from mature blood cells of thrombosis models in vitro and in vivo because of targeting RNA, while simultaneously image microviscosity changes in these cells. So, dABp-3 as an analytical tool holds considerable promise for bioimaging and monitoring of microviscosity changes in complex biological systems.
机译:活细胞的微粘度变化对RNA的扩散和移动能力具有深远的影响,更严重的是,它可以修饰活细胞中的RNA功能。荧光转子的荧光响应不同的环境粘度,在生物系统中的粘度成像方面具有巨大的潜力。尽管已经报道了许多荧光转头的粘度,但很少报道具有超灵敏性的荧光转头,用于测定微粘度(<10 cP)。在这里,我们报道了一种基于核苷的双光子荧光转子(dABp-3),它​​可以选择性地和超灵敏地首次成像活细胞RNA区域中的微粘度。选择2'-脱氧腺苷作为电子供体,以允许能量通过炔键传递至受体(典型的硼二吡咯亚甲基部分)。另一个亮点是dABp-3基于2'-脱氧腺苷,这导致其对RNA的识别能力。 dABp-3具有超敏性,在双模式双光子比率模式和荧光寿命模式下,对活细胞RNA区域中的微范围粘度(1.8-6.0 cP)提供了变化的线性响应。在筛选和优化之后,有利的是,由于靶向RNA,dABp-3可用于在体外和体内从血栓形成模型的成熟血细胞中筛选网状细胞,同时对这些细胞中的微粘度变化进行成像。因此,dABp-3作为分析工具在生物成像和监测复杂生物系统中微粘度变化方面具有广阔的前景。

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