1. Super-Resolution Imaging of the Golgi in Live Cells with a Bioorthogonal Ceramide Probe
- Author
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Hideo Takakura, Edward S. Allgeyer, Joerg Bewersdorf, Felix Rivera-Molina, Derek Toomre, Alanna Schepartz, Alexander D. Thompson, and Roman S. Erdmann
- Subjects
Ceramide ,Cell Survival ,1.1 Normal biological development and functioning ,Confocal ,Golgi Apparatus ,Ceramides ,Fluorescence ,Article ,Catalysis ,chemistry.chemical_compound ,symbols.namesake ,Underpinning research ,fluorophores ,Humans ,Coloring Agents ,Microscopy ,Microscopy, Confocal ,Endoplasmic reticulum ,Organic Chemistry ,STED microscopy ,General Chemistry ,Golgi apparatus ,bioorthogonal chemistry ,3. Good health ,STED ,Membrane ,Microscopy, Fluorescence ,chemistry ,Biochemistry ,membranes ,Hela Cells ,click chemistry ,Chemical Sciences ,Click chemistry ,Biophysics ,symbols ,lipids (amino acids, peptides, and proteins) ,Generic health relevance ,Bioorthogonal chemistry ,HeLa Cells - 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 non-toxic 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.
- Published
- 2014
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