ArticleJournal of lipid research2016
A highly sensitive protocol for microscopy of alkyne lipids and fluorescently tagged or immunostained proteins.
Article in Journal of lipid research, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 26 citations in OpenAlex.
- Bioconjugation Strategies for Revealing the Roles of Glycerophospholipids in Cells.Chembiochem : a European journal of chemical biology · 2026Review
- 1-deoxysphingolipids dysregulate membrane properties and cargo trafficking in the early secretory pathway.Cell chemical biology · 2026Article
- Analytical Techniques for Single-Cell Biochemical Assays of Lipids.Annual review of biomedical engineering · 2023Review
- Chemical Probes to Control and Visualize Lipid Metabolism in the Brain.Accounts of chemical research · 2022Review
- Combining Isoprenoid Probes with Antibody Markers for Mass Cytometric Analysis of Prenylation in Single Cells.Analytical chemistry · 2022Article
- PUFA-DerivedACS chemical biology · 2022Article
- Tracing Lipid Metabolism by Alkyne Lipids and Mass Spectrometry: The State of the Art.Frontiers in molecular biosciences · 2022Review
- A Not-So-Ancient Grease History: Click Chemistry and Protein Lipid Modifications.Chemical reviews · 2021Review
- Plasmon-assisted click chemistry at low temperature: an inverse temperature effect on the reaction rate.Chemical science · 2021Article
- Clickable Vitamins as a New Tool to Track Vitamin A and Retinoic Acid in Immune Cells.Frontiers in immunology · 2021Article
- Glycolysis regulates Hedgehog signalling via the plasma membrane potential.The EMBO journal · 2020Article
- Multiplexed and single cell tracing of lipid metabolism.Nature methods · 2019Article
- Double Nitroxide Labeling by Copper-Catalyzed Azide-Alkyne Cycloadditions with Noncanonical Amino Acids for Electron Paramagnetic Resonance Spectroscopy.ACS chemical biology · 2019Article
- Localization of 1-deoxysphingolipids to mitochondria induces mitochondrial dysfunction.Journal of lipid research · 2017Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The demand to study the cellular localization of specific lipids has led to recent advances in lipid probes and microscopy. Alkyne lipids bear a small, noninterfering tag and can be detected upon click reaction with an azide-coupled reporter. Fluorescent alkyne lipid imaging crucially depends on appropriate azide reporters and labeling protocols that allow for an efficient click reaction and therefore a sensitive detection. We synthesized several azide reporters with different spacer components and tested their suitability for alkyne lipid imaging in fixed cells. The implementation of a copper-chelating picolyl moiety into fluorescent or biotin-based azide reagents strongly increased the sensitivity of the imaging routine. We demonstrate the applicability and evaluate the performance of this approach using different lipid classes and experimental setups. As azide picolyl reporters allow for reduced copper catalyst concentrations, they also enable coimaging of alkyne lipids with multiple fluorescent proteins including enhanced green fluorescent protein. Alternatively, and as we also show, microscopy of alkyne lipids can be combined with protein detection by immunocytochemistry. In summary, we present a robust, sensitive, and highly versatile protocol for the labeling of alkyne lipids with azide-coupled reporters for fluorescence microscopy that can be combined with different protein detection and imaging techniques.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.