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Revised Diffusion Law Permits Quantitative Nanoscale Characterization of Membrane Organization
J. Heyrovský Institute of Physical Chemistry of the Czech Academy of Sciences, Dolejškova 3, Prague 182 23, Czech Republic; Faculty of Mathematics and Physics, Charles University, Ke Karlovu 5, Prague 121 16, Czech Republic.
J. Heyrovský Institute of Physical Chemistry of the Czech Academy of Sciences, Dolejškova 3, Prague 182 23, Czech Republic; Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Hlavova 8, Prague 128 40, Czech Republic.
KTH, School of Engineering Sciences (SCI), Applied Physics. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0002-5584-9170
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Rusian Academy of Science, Moscow 117997, Russia.
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2025 (English)In: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 97, no 22, p. 11478-11485Article in journal (Refereed) Published
Abstract [en]

The formation of functional nanoscopic domains is an inherent property of plasma membranes. Stimulated emission depletion combined with fluorescence correlation spectroscopy (STED-FCS) has been previously used to identify such domains; however, the information obtained by STED-FCS has been limited to the presence of such domains while crucial parameters have not been accessible, such as size (R d), the fraction of occupied membrane surface (f), in-membrane lipid diffusion inside (D in) and outside (D out) the nanodomains as well as their self-diffusion (D d). Here, we introduce a quantitative approach based on a revised interpretation of the diffusion law. By analyzing experimentally recorded STED-FCS diffusion law plots using a comprehensive library of simulated diffusion law plots, we extract these five parameters from STED-FCS data. That approach is verified on ganglioside nanodomains in giant unilamellar vesicles, validating the Saffman-Delbruck assumption for D d. STED-FCS data in both plasma membranes of living PtK2 cells and giant plasma membrane vesicles are examined, and a quantitative framework for molecular diffusion modes in biological membranes is presented.

Place, publisher, year, edition, pages
American Chemical Society (ACS) , 2025. Vol. 97, no 22, p. 11478-11485
National Category
Biophysics
Identifiers
URN: urn:nbn:se:kth:diva-367929DOI: 10.1021/acs.analchem.5c00021ISI: 001498667900001PubMedID: 40437882Scopus ID: 2-s2.0-105006812288OAI: oai:DiVA.org:kth-367929DiVA, id: diva2:1987008
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QC 20260120

Available from: 2025-08-04 Created: 2025-08-04 Last updated: 2026-01-20Bibliographically approved

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Blom, Hans

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