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Super‐Resolution Imaging of the Golgi in Live Cells with a Bioorthogonal Ceramide Probe
- Erdmann, Roman S;
- Takakura, Hideo;
- Thompson, Alexander D;
- Rivera‐Molina, Felix;
- Allgeyer, Edward S;
- Bewersdorf, Joerg;
- Toomre, Derek;
- Schepartz, Alanna
Published Web Location
https://doi.org/10.1002/anie.201403349Abstract
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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