Super‐Resolution Imaging of the Golgi in Live Cells with a Bioorthogonal Ceramide Probe |
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| Authors: | Dr Roman S Erdmann Dr Hideo Takakura Alexander D Thompson Felix Rivera‐Molina Dr Edward S Allgeyer Prof?Dr Joerg Bewersdorf Prof?Dr Derek Toomre Prof?Dr Alanna Schepartz |
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| Affiliation: | 1. R.S.E. and H.T. contributed equally to this work.;2. Department of Chemistry, Yale University, 225 Prospect Street, New Haven CT 06511 (USA);3. Department of Cell Biology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520 (USA) |
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| 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. |
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| Keywords: | bioorthogonal chemistry click chemistry fluorophores membranes STED |
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