title: Super-resolution imaging of the Golgi in live cells with a bioorthogonal ceramide probe.
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.
authors: Department 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.
authors: Roman S
authors: RS
authors: Erdmann
authors: Hideo
authors: H
authors: Takakura
authors: Alexander D
authors: AD
authors: Thompson
authors: Felix
authors: F
authors: Rivera-Molina
authors: Edward S
authors: ES
authors: Allgeyer
authors: Joerg
authors: J
authors: Bewersdorf
authors: Derek
authors: D
authors: Toomre
authors: Alanna
authors: A
authors: Schepartz
mesh_terms: D002470:Cell Survival; D002518:Ceramides / Q000737:chemistry*; D004396:Coloring Agents / Q000032:analysis* / Q000737:chemistry*; D006056:Golgi Apparatus / Q000201:enzymology / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence
chemical_list: D002518:Ceramides; D004396:Coloring Agents
keywords: STED; bioorthogonal chemistry; click chemistry; fluorophores; membranes