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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??M 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.

SUBMITTER: Erdmann RS

PROVIDER: S-EPMC4593319 | biostudies-literature | 2014 Sep

REPOSITORIES: biostudies-literature

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

Erdmann Roman S RS Takakura Hideo H Thompson Alexander D AD Rivera-Molina Felix F Allgeyer Edward S ES Bewersdorf Joerg J Toomre Derek D Schepartz Alanna A

Angewandte Chemie (International ed. in English) 20140731 38

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 ...[more]

PMID: 25081303

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