Design and Preparation of Lifetime-Based Dual Fluorescent/Phosphorescent Sensor of pH and Oxygen and its Exploration in Model Physiological Solutions and Cells

IF 4.4 4区 医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Vadim Baigildin, Julia Shakirova, Nina Zharskaia, Elena Ivanova, Sergey Silonov, Viktor Sokolov, Sergey Tunik
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Abstract

In the present report, a novel dual pH-O2 sensor based on covalent conjugate of rhodamine 6G and cyclometalated iridium complex with poly(vinylpyrrolidone-block-vinyltetrazole) copolymer is reported. In model physiological solutions the sensor chromophores display independent phosphorescent and fluorescent lifetime responses onto variations in oxygen concentration and pH, respectively. Colocalization studies on Chinese hamster ovary cells demonstrate the preferential localization in endosomes and lysosomes. The fluorescent lifetime imaging microscopy-phosphorescent lifetime imaging microscopy (FLIM-PLIM) experiments show that the phosphorescent O2 sensor provides unambiguous information onto hypoxia versus normoxia cell status as well as semi-quantitative data on the oxygen concentration in cells in between these two states. However, the results of FLIM measurements indicate that dynamic lifetime interval of the sensor (≈0.5 ns between pH values 5.0 and 8.0) is insufficient even for qualitative estimation of pH in living cells because half-width of lifetime distribution in the studied samples is higher than the sensor dynamic interval. Nevertheless, the variations in rhodamine emission intensity are much higher and allow rough discrimination of acidic and neutral cell conditions. Thus, the results of this study indicate that the suggested approach to the design of dual pH-O2 sensors makes possible to prepare the biocompatible and water-soluble conjugate with fast cellular uptake.

基于生命周期的 pH 和氧气双荧光/磷光传感器的设计和制备及其在模型生理溶液和细胞中的应用探索。
本报告介绍了一种新型 pH-O2 双传感器,它基于罗丹明 6G 和环甲基铱络合物与聚乙烯吡咯烷酮-块状-乙烯基四唑共聚物的共价共轭物。在模型生理溶液中,传感器发色团分别对氧浓度和 pH 值的变化显示出独立的磷光和荧光寿命响应。对中国仓鼠卵巢细胞进行的共定位研究表明,传感器优先定位在内膜和溶酶体中。荧光寿命成像显微镜-磷光寿命成像显微镜(FLIM-PLIM)实验表明,磷光 O2 传感器可提供缺氧与正常缺氧细胞状态的明确信息,以及介于这两种状态之间的细胞中氧浓度的半定量数据。然而,FLIM 测量结果表明,传感器的动态寿命间隔(pH 值 5.0 和 8.0 之间≈0.5 毫微秒)甚至不足以对活细胞中的 pH 值进行定性估计,因为所研究样品中的寿命分布半宽度高于传感器的动态间隔。不过,罗丹明发射强度的变化要大得多,可以粗略区分酸性和中性细胞条件。因此,这项研究的结果表明,所建议的 pH-O2 双传感器设计方法可以制备出生物相容性好、可溶于水且能被细胞快速吸收的共轭物。
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来源期刊
Macromolecular bioscience
Macromolecular bioscience 生物-材料科学:生物材料
CiteScore
7.90
自引率
2.20%
发文量
211
审稿时长
1.5 months
期刊介绍: Macromolecular Bioscience is a leading journal at the intersection of polymer and materials sciences with life science and medicine. With an Impact Factor of 2.895 (2018 Journal Impact Factor, Journal Citation Reports (Clarivate Analytics, 2019)), it is currently ranked among the top biomaterials and polymer journals. Macromolecular Bioscience offers an attractive mixture of high-quality Reviews, Feature Articles, Communications, and Full Papers. With average reviewing times below 30 days, publication times of 2.5 months and listing in all major indices, including Medline, Macromolecular Bioscience is the journal of choice for your best contributions at the intersection of polymer and life sciences.
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