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Super‐Resolution Imaging of the Golgi in Live Cells with a Bioorthogonal Ceramide Probe
Author(s) -
Erdmann Roman S.,
Takakura Hideo,
Thompson Alexander D.,
RiveraMolina Felix,
Allgeyer Edward S.,
Bewersdorf Joerg,
Toomre Derek,
Schepartz Alanna
Publication year - 2014
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201403349
Subject(s) - golgi apparatus , bioorthogonal chemistry , sted microscopy , endoplasmic reticulum , ceramide , microbiology and biotechnology , chemistry , live cell imaging , tetrazine , organelle , biophysics , nanotechnology , biology , biochemistry , cell , materials science , superresolution , click chemistry , combinatorial chemistry , apoptosis , image (mathematics) , organic chemistry , artificial intelligence , computer science
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 μ 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.