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利用铱-炔探针对活细胞内合成蛋白质进行光致发光寿命成像。

Photoluminescence Lifetime Imaging of Synthesized Proteins in Living Cells Using an Iridium-Alkyne Probe.

机构信息

Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing, 100084, China.

Key Lab of Organic Optoelectronics and Molecular Engineering of Ministry of Education, Department of Chemistry, Tsinghua University, Beijing, 100084, China.

出版信息

Angew Chem Int Ed Engl. 2017 Nov 20;56(47):14928-14932. doi: 10.1002/anie.201708566. Epub 2017 Oct 13.

Abstract

Designing probes for real-time imaging of dynamic processes in living cells is a continuous challenge. Herein, a novel near-infrared (NIR) photoluminescence probe having a long lifetime was exploited for photoluminescence lifetime imaging (PLIM) using an iridium-alkyne complex. This probe offers the benefits of deep-red to NIR emission, a long Stokes shift, excellent cell penetration, low cytotoxicity, and good resistance to photobleaching. This example is the first PLIM probe applicable to the click reaction of copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) with remarkable lifetime shifts of 414 ns, before and after click reaction. The approach fully eliminates the background interference and distinguishes the reacted probes from the unreacted probes, thus enabling the wash-free imaging of the newly synthesized proteins within single living cells. Based on the unique properties of the iridium complexes, it is anticipated to have applications for imaging other processes within living cells.

摘要

设计用于实时成像活细胞中动态过程的探针是一个持续的挑战。在此,利用铱-炔烃配合物开发了一种新型的近红外(NIR)光致发光探针,用于光致发光寿命成像(PLIM)。该探针具有深红色到 NIR 发射、大斯托克斯位移、出色的细胞穿透性、低细胞毒性和良好的抗光漂白性等优点。该示例是第一个适用于铜(I)催化的叠氮化物-炔烃环加成(CuAAC)点击反应的 PLIM 探针,在点击反应前后具有 414ns 的显著寿命变化。该方法完全消除了背景干扰,并区分了反应探针和未反应探针,从而能够在单个活细胞内无冲洗地对新合成的蛋白质进行成像。基于铱配合物的独特性质,预计其在活细胞内的其他过程成像中具有应用前景。

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