Revised Diffusion Law Permits Quantitative Nanoscale Characterization of Membrane OrganizationShow others and affiliations
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
Note
QC 20260120
2025-08-042025-08-042026-01-20Bibliographically approved