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Angew Chem Int Ed Engl. 2014 Sep 15;53(38):10242-6. doi: 10.1002/anie.201403349. Epub 2014 Jul 31.

Super-resolution imaging of the Golgi in live cells with a bioorthogonal ceramide probe.

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  • 1Department of Chemistry, Yale University, 225 Prospect Street, New Haven CT 06511 (USA); Department of Cell Biology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520 (USA); R.S.E. and H.T. contributed equally to this work.

Abstract

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 nontoxic 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.

© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

KEYWORDS:

STED; bioorthogonal chemistry; click chemistry; fluorophores; membranes

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