Super‐resolution microscopy using a bioorthogonal‐based cholesterol probe provides unprecedented capabilities for imaging nanoscale lipid heterogeneity in living cells
File(s)
Published version
Author(s)
Type
Journal Article
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-N3) 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-N3 probe mimics cholesterol in synthetic and cellular membranes without perturbation. When combined with live-cell super-resolution microscopy, chol-N3 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.
Date Issued
2021-09-15
Date Acceptance
2021-07-01
Citation
Small Methods, 2021, 5 (9), pp.1-15
ISSN
2366-9608
Publisher
Wiley
Start Page
1
End Page
15
Journal / Book Title
Small Methods
Volume
5
Issue
9
Copyright Statement
© 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.
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.
License URL
Sponsor
Bill and Melinda Gates Foundation
Biotechnology and Biological Sciences Research Council
Biotechnology and Biological Sciences Research Council
Identifier
https://onlinelibrary.wiley.com/doi/10.1002/smtd.202100430
Grant Number
INV-016631
BB/V019791/1
BB/V019791/1
Subjects
Science & Technology
Physical Sciences
Technology
Chemistry, Physical
Nanoscience & Nanotechnology
Materials Science, Multidisciplinary
Chemistry
Science & Technology - Other Topics
Materials Science
bioorthogonal reactions
cholesterol
lipid raft
membranes
nanoprobes
nanoscale lipid heterogeneity
super-resolution microscopy
GENERALIZED POLARIZATION
RAFTS
MEMBRANES
ORGANIZATION
TRANSITION
VESICLES
DOMAINS
ORDER
Publication Status
Published
Date Publish Online
2021-07-29