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荧光成像法通过 3-(dansylamino)苯硼酸与细胞表面唾液酸的竞争结合来原位检测细胞表面唾液酸。

Fluorescence imaging for in situ detection of cell surface sialic acid by competitive binding of 3-(dansylamino)phenylboronic acid.

机构信息

State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210093, PR China.

State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210093, PR China.

出版信息

Anal Chim Acta. 2015 Sep 24;894:85-90. doi: 10.1016/j.aca.2015.08.054. Epub 2015 Sep 3.

Abstract

Sialic acid (SA) usually locates at the terminal position of the sugar chains on cell membranes, and its expression level is closely associated with cancer. Here polysialic acid (PSA) embedded gold nanoparticles (AuNPs) were prepared and functionalized with fluorescent 3-(dansylamino)phenylboronic acid (DAPB) for in situ imaging and detection of cell surface SA. The fluorescence resonance energy transfer (FRET) from DAPB to AuNPs quenched the fluorescence of DAPB. In the presence of additional SA or SA-abundant cells, the competitive binding of DAPB with SA and PSA led to the release of the assembled DAPB from the surface of PSA-embedded AuNPs, resulting in fluorescence of DAPB on SA-abundant cell surface. The proposed methods realized the in situ imaging and monitoring of cell surface SA, and could also be applied to the quantification of cell number and the amounts of cell surface SA. This work not only proposed a convenient visualization method for the analysis of SA on cell membranes, but also provided a potential tool for accelerating the elucidation of the basic role of SA in various biological processes and development of anti-cancer therapies.

摘要

唾液酸(SA)通常位于细胞膜糖链的末端位置,其表达水平与癌症密切相关。本研究制备了嵌入聚唾液酸(PSA)的金纳米粒子(AuNPs),并用荧光 3-(丹磺酰基氨基)苯硼酸(DAPB)进行功能化,用于细胞表面 SA 的原位成像和检测。DAPB 到 AuNPs 的荧光共振能量转移(FRET)猝灭了 DAPB 的荧光。在存在额外的 SA 或富含 SA 的细胞的情况下,DAPB 与 SA 和 PSA 的竞争结合导致组装的 DAPB 从 PSA 嵌入的 AuNPs 表面释放,从而在富含 SA 的细胞表面上产生 DAPB 的荧光。所提出的方法实现了细胞表面 SA 的原位成像和监测,并且还可以应用于细胞数量和细胞表面 SA 量的定量。这项工作不仅提出了一种用于分析细胞膜上 SA 的方便可视化方法,而且还为加速阐明 SA 在各种生物过程中的基本作用和开发抗癌疗法提供了一种潜在的工具。

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