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Updated: Apr 26, 2026
Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass
Published on: August 10, 2017
Super-resolution imaging of the Golgi in live cells with a bioorthogonal ceramide probe
Roman S Erdmann1, Hideo Takakura, Alexander D Thompson
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 microM 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.
