ArticleBiophysical journal2015
Self-calibrated line-scan STED-FCS to quantify lipid dynamics in model and cell membranes.
Article in Biophysical journal, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Super-resolution microscopy to study membrane nanodomains and transport mechanisms in the plasma membrane.Frontiers in molecular biosciences · 2024Review
- Phosphorylation of tyrosine 90 in SH3 domain is a new regulatory switch controlling Src kinase.eLife · 2023Article
- Nanowaveguide-illuminated fluorescence correlation spectroscopy for single molecule studies.AIP advances · 2021Article
- Article
- z-STED Imaging and Spectroscopy to Investigate Nanoscale Membrane Structure and Dynamics.Biophysical journal · 2020Article
- Clustering of the ζ-Chain Can Initiate T Cell Receptor Signaling.International journal of molecular sciences · 2020Article
- Variations in Plasma Membrane Topography Can Explain Heterogenous Diffusion Coefficients Obtained by Fluorescence Correlation Spectroscopy.Frontiers in cell and developmental biology · 2020Article
- HIV-1 Gag specifically restricts PI(4,5)P2 and cholesterol mobility in living cells creating a nanodomain platform for virus assembly.Science advances · 2019Article
- Measuring nanoscale diffusion dynamics in cellular membranes with super-resolution STED-FCS.Nature protocols · 2019Article
- Article
- Nanoscale Spatiotemporal Diffusion Modes Measured by Simultaneous Confocal and Stimulated Emission Depletion Nanoscopy Imaging.Nano letters · 2018Article
- Super-resolution fluorescence microscopy studies of human immunodeficiency virus.Retrovirology · 2018Review
- Micro-pharmacokinetics: Quantifying local drug concentration at live cell membranes.Scientific reports · 2018Article
- Envelope glycoprotein mobility on HIV-1 particles depends on the virus maturation state.Nature communications · 2017Article
- Measurement of nanoscale three-dimensional diffusion in the interior of living cells by STED-FCS.Nature communications · 2017Article
- An intermolecular FRET sensor detects the dynamics of T cell receptor clustering.Nature communications · 2017Article
- Raft-based sphingomyelin interactions revealed by new fluorescent sphingomyelin analogs.The Journal of cell biology · 2017Article
- A FRET sensor enables quantitative measurements of membrane charges in live cells.Nature biotechnology · 2017Article
- Convergence of lateral dynamic measurements in the plasma membrane of live cells from single particle tracking and STED-FCS.Journal of physics D: Applied physics · 2017Review
- Biologists Wanted: New Fluorescence Fluctuation Tools for Cell Biology.Biophysical journal · 2016Article
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Only a limited number of noninvasive techniques are available to directly measure the dynamic behavior of lipids in model and cell membranes. Here, we explored whether a commercial instrument could be used for fluorescence correlation spectroscopy (FCS) under pulsed stimulated emission depletion (STED). To overcome issues with photobleaching and poor distinction between confocal and STED signals, we implemented resonant line-scan STED with filtered FCS, which has the additional benefit of autocalibrating the dimensions of the point-spread function and obtaining spatially resolved molecular mobility at subdiffraction resolution. With supported lipid bilayers, we achieved a detection spot radius of 40 nm, although at the expense of decreased molecular brightness. We also used this approach to map the dynamics of Atto646N-labeled sphingomyelin and phosphatidylethanolamine in the plasma membrane. Despite the reliability of the method and the demonstration that photobleaching and the photophysical properties of the dye did not influence diffusion measurements, we found great heterogeneities even within one cell. For both lipids, regions of high local density correlated with slow molecular diffusion, indicating trapping of Atto646N-labeled lipids. Future studies with new dyes are needed to reveal the origin of the trapping.
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