超分辨显微镜（STORM）合成和细胞内示踪表面功能化磷酸钙纳米颗粒

广州微视光学科技有限公司

2023/11/07 16:35

通过铜催化的叠氮化物-炔烃环加成反应（SPAA）和应变促进的叠氮化物-炔烃环加成反应（点击化学反应）制备了具有共价连接的荧光染料分子的磷酸钙纳米粒子。比较并量化了两次点击反应的反应动力学和标记度。对于氧化敏感性分子，由于反应条件较温和且不存在铜（II），因此首选SPAAC方法。高荧光AF-488标记的纳米颗粒用于超分辨率荧光显微镜，即随机光学重建显微镜（STORM）。通过这种方法，在细胞以20–30 nm的分辨率摄取后，在HeLa细胞内部的溶酶体中观察到了单个纳米颗粒（直径约60 nm）。

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Synthesis and intracellular tracing surface-functionalized calcium phosphate nanoparticles by super-resolution microscopy (STORM)

Abstract Calcium phosphate nanoparticles with covalently attached fluorescent dye molecules were prepared by copper-catalysed azide-alkyne cycloaddition (CuAAC) and by strain-promoted azide-alkyne cycloaddition (SPAAC), i.e. click chemistry. The reaction kinetics and labeling degrees of the two click reactions were compared and quantified. For oxidation-sensitive molecules, the SPAAC method is preferable due to the milder reaction conditions and the absence of copper(II). The highly fluorescent AF-488-labeled nanoparticles were applied for super-resolution fluorescence microscopy, i.e. stochastic optical reconstruction microscopy (STORM). By this method, individual nanoparticles (diameter about 60 nm) were observed in endolysosomes inside HeLa cells after cellular uptake at a resolution of 20–30 nm.

原文链接：超分辨显微镜（STORM）合成和细胞内示踪表面功能化磷酸钙纳米颗粒 - ScienceDirect

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