Super-Resolution Microscopy Using a Bioorthogonal-Based Cholesterol Probe Provides Unprecedented Capabilities for Imaging Nanoscale Lipid Heterogeneity in Living Cells
dc.contributor.author | Lorizate Nogales, Maier | |
dc.contributor.author | Terrones Urio, Oihana | |
dc.contributor.author | Nieto Garai, Jon Ander | |
dc.contributor.author | Rojo Bartolomé, Iratxe | |
dc.contributor.author | Ciceri, Dalila | |
dc.contributor.author | Morana, Ornella | |
dc.contributor.author | Olazar Intxausti, June | |
dc.contributor.author | Arboleya, Aroa | |
dc.contributor.author | Martin, Alexia | |
dc.contributor.author | Szynkiewicz, Marta | |
dc.contributor.author | Calleja Felipe, María | |
dc.contributor.author | Bernardino de la Serna, Jorge | |
dc.contributor.author | Contreras, F. Xabier | |
dc.date.accessioned | 2021-09-21T12:34:20Z | |
dc.date.available | 2021-09-21T12:34:20Z | |
dc.date.issued | 2021-09 | |
dc.identifier.citation | Small Methods 5(9) : (2021) // Article ID 2100430 | es_ES |
dc.identifier.issn | 2366-9608 | |
dc.identifier.uri | http://hdl.handle.net/10810/53119 | |
dc.description.abstract | Despite more than 20 years of work since the lipid raft concept was proposed, the existence of these nanostructures remains highly controversial due to the lack of noninvasive methods to investigate their native nanorganization in living unperturbed cells. There is an unmet need for probes for direct imaging of nanoscale membrane dynamics with high spatial and temporal resolution in living cells. In this paper, a bioorthogonal-based cholesterol probe (chol-N-3) is developed that, combined with nanoscopy, becomes a new powerful method for direct visualization and characterization of lipid raft at unprecedented resolution in living cells. The chol-N-3 probe mimics cholesterol in synthetic and cellular membranes without perturbation. When combined with live-cell super-resolution microscopy, chol-N-3 demonstrates the existence of cholesterol-rich nanodomains of <50 nm at the plasma membrane of resting living cells. Using this tool, the lipid membrane structure of such subdiffraction limit domains is identified, and the nanoscale spatiotemporal organization of cholesterol in the plasma membrane of living cells reveals multiple cholesterol diffusion modes at different spatial localizations. Finally, imaging across thick organ samples outlines the potential of this new method to address essential biological questions that were previously beyond reach. | es_ES |
dc.description.sponsorship | M.L., O.T., and J.A.N.-G. contributed equally to this work. This work was supported by grants from the Spanish Ministry of Science Innovation and Universities, (Grant No. BFU-2015-68981-P) and the Basque Government (Grant No. IT1264-19) to F.-X.C. and M.L.. The authors thank J. M. Gonzalez Manas and Sergio Perez Acebron for helful comments on the manuscript. The authors thank the technical and human support provided by the analytical and high-resolution microscopy facility (SGIker) of UPV/EHU and European funding (ERDF and ESF). J.B.d.l.S. acknowledges funding from the Bill and Melinda Gates Foundation and the BBSRC (Grant Nos. INV-016631 and BB/V019791/1, respectively). This work was supported in part by the Fundacion Biofisica Bizkaia (FBB) and the Basque Excellence Research Centre (BERC) program of the Basque Government. J.A.N.-G. was supported by a FI predoctoral fellowship from the Basque Government and currently by FBB. Document | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | Wiley | es_ES |
dc.relation | info:eu-repo/grantAgreement/MICINN/BFU-2015-68981-P | es_ES |
dc.rights | info:eu-repo/semantics/openAccess | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by/3.0/es/ | * |
dc.subject | bioorthogonal reactions | es_ES |
dc.subject | cholesterol | es_ES |
dc.subject | lipid raft | es_ES |
dc.subject | membranes | es_ES |
dc.subject | nanoprobes | es_ES |
dc.subject | nanoscale lipid heterogeneity | es_ES |
dc.subject | super-resolution microscopy | es_ES |
dc.subject | generalized polarization | es_ES |
dc.subject | rafts | es_ES |
dc.subject | organization | es_ES |
dc.subject | transition | es_ES |
dc.subject | vesicles | es_ES |
dc.subject | domains | es_ES |
dc.subject | order | es_ES |
dc.title | Super-Resolution Microscopy Using a Bioorthogonal-Based Cholesterol Probe Provides Unprecedented Capabilities for Imaging Nanoscale Lipid Heterogeneity in Living Cells | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.rights.holder | 2021 The Authors. Small Methods published by Wiley-VCH GmbH. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. | es_ES |
dc.rights.holder | Atribución 3.0 España | * |
dc.relation.publisherversion | https://onlinelibrary.wiley.com/doi/10.1002/smtd.202100430 | es_ES |
dc.identifier.doi | 10.1002/smtd.202100430 | |
dc.departamentoes | Bioquímica y biología molecular | es_ES |
dc.departamentoeu | Biokimika eta biologia molekularra | es_ES |
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Except where otherwise noted, this item's license is described as 2021 The Authors. Small Methods published by Wiley-VCH GmbH. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in
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