Super-resolution imaging of the Golgi in live cells with a bioorthogonal ceramide probe.
Super-resolution imaging of the Golgi in live cells with a bioorthogonal ceramide probe.
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DOI：
10.1002/anie.201403349
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发表时间：
2014-09-15
期刊：
影响因子：
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通讯作者：
Schepartz A
中科院分区：
文献类型：
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作者：
Erdmann RS;Takakura H;Thompson AD;Rivera-Molina F;Allgeyer ES;Bewersdorf J;Toomre D;Schepartz A
我们报告了一种基于脂质的策略，以超分辨率在活细胞中可视化高尔基体结构和动态。该方法基于两种新型试剂：含反式环辛烯的神经酰胺脂质（Cer-TCO）和高反应性四嗪标记的近红外染料（SiR-Tz）。这些试剂通过极其快速的“四嗪点击”反应组装成Cer-SiR，Cer-SiR是一种高度光稳定的“活体染料”，能够通过3D共聚焦和STED显微镜对高尔基体进行长时间的活细胞成像。Cer-SiR在高达2 μM的浓度下是无毒的，并且不会干扰高尔基体驻留酶的移动性或从内质网通过高尔基体到质膜的货物运输。
We report a lipid-based strategy to visualize Golgi structure and dynamics at super-resolution in live cells. The method is based on two novel reagents: a trans-cyclooctene-containing ceramide lipid (Cer-TCO) and a highly reactive, tetrazine-tagged near-IR dye (SiR-Tz). These reagents assemble via an extremely rapid ‘tetrazine-click’ reaction into Cer-SiR, a highly photostable ‘vital dye’ that enables prolonged live cell imaging of the Golgi apparatus by 3D confocal and STED microscopy. Cer-SiR is non-toxic at concentrations as high as 2 μM and does not perturb the mobility of Golgi-resident enzymes or the traffic of cargo from the endoplasmic reticulum through the Golgi and to the plasma membrane.