| Literature DB >> 28924659 |
Roman S Erdmann1,2, Derek Toomre1, Alanna Schepartz3,4.
Abstract
Long time-lapse super-resolution imaging in live cells requires a labeling strategy that combines a bright, photostable fluorophore with a high-density localization probe. Lipids are ideal high-density localization probes, as they are >100 times more abundant than most membrane-bound proteins and simultaneously demark the boundaries of cellular organelles. Here, we describe Cer-SiR, a two-component, high-density lipid probe that is exceptionally photostable. Cer-SiR is generated in cells via a bioorthogonal reaction of two components: a ceramide lipid tagged with trans-cyclooctene (Cer-TCO) and a reactive, photostable Si-rhodamine dye (SiR-Tz). These components assemble within the Golgi apparatus of live cells to form Cer-SiR. Cer-SiR is benign to cellular function, localizes within the Golgi at a high density, and is sufficiently photostable to enable visualization of Golgi structure and dynamics by 3D confocal or long time-lapse STED microscopy.Entities:
Keywords: Bioorthogonal chemistry; Click chemistry; Fluorophores; Inverse electron demand Diels-Alder reaction; Membranes; Super-resolution microscopy
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Year: 2017 PMID： 28924659 PMCID： PMC6146391 DOI： 10.1007/978-1-4939-7265-4_6
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745