High‐Speed Interferometric Scattering Tracking Microscopy of Compartmentalized Lipid Diffusion in Living Cells
Year: 2025
Type: article
Abstract: Lateral diffusion measurements have been ‐used to infer information about the nano‐organization of membranes. We employed interferometric scattering (ISCAT) microscopy at an acquisition rate of 2 kHz to revisit the diffusion dynamics of a phospholipid analog on the plasma membrane of Ptk2 cells. The ISCAT trajectory data are analyzed with an unbiased, statistics‐driven pipeline to identify the most likely diffusion mode from a set of plausible diffusion modes. At the ensemble average level, the data are best described as transient compartmentalized diffusion with an average compartment size of 100–110 nm, transient confinement time of 8–10 ms, intracompartmental diffusion coefficient of 0.7–0.9 μm 2 s −1 , and intercompartmental diffusion coefficient of 0.3–0.4 μm 2 s −1 . The same analysis applied at the single‐trajectory level identifies a complex variety of diffusion modes with 7–8% free, 13–14% confined, 40% transient compartmentalized, and 40% anomalous diffusion. Measurements with larger (Ø40 nm) as compared to smaller (Ø20 nm) gold nanoparticles are found to influence the diffusion rate and confinement strength, but not the underlying lipid diffusion modes. Using Monte Carlo simulations, these experimental results are explored in the wider context of relevant literature. This analysis paints a unifying picture of lipid diffusion on mammalian cell membranes transcending differences between experimental techniques.
Source: ChemPhysChem
Institutions: University of Vienna, Leibniz Institute of Photonic Technology, John Radcliffe Hospital, Max Perutz Labs, University of Oxford, MRC Weatherall Institute of Molecular Medicine, MRC Human Immunology Unit, Friedrich Schiller University Jena, Åbo Akademi University, University of Turku, Kennedy Institute of Rheumatology
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $4,550
Funders: Wellcome Trust, Friedrich-Schiller-Universität Jena, Freistaat Thüringen, University of Oxford, Deutsche Forschungsgemeinschaft, Bundesministerium für Bildung und Forschung, Thüringer Ministerium für Wirtschaft, Wissenschaft und Digitale Gesellschaft, Medical Research Council, Engineering and Physical Sciences Research Council, Biotechnology and Biological Sciences Research Council, Leibniz-Gemeinschaft, EPA Cephalosporin Fund
Awards: G1215500115, G1515183235, G1781628337, G1809803311, G1885323389, MICA: Nanoscopy Oxford (NanO): Novel Super-Resolution Imaging Applied to Biomedical Sciences, G2205214540, G281949974, G2967549892, SFB 1278: Polymerbasierte NanopartikelBibliotheken für die Entwicklung zielgerichteter antiinflammatorischer Strategien, G3056851145, G3065514043, G3423061657, EPSRC and MRC Centre for Doctoral Training in Biomedical Imaging, Verbundprojekt: Hochparalleles Profiling der Wirtsantwort auf lebensbedrohliche Infektionen (LPI-BT4) - Teilvorhaben: Photonische Technologien für hochparalleles Profiling der Wirtsantwort auf lebensbedrohliche Infektionen, Verbundprojekt: Innovative molekulare und biochemische Assays für Schnelldiagnostik, Drug Development und neue Therapiekonzepte (LPI-BT5) - Teilvorhaben: Entwicklung molekularer Techniken, Probenvorbereitungsstrategien und photonischer Detektionsmethoden zum Nachweis und Charakterisierung von Mikroorganismen, G4537655822, G4632173870, G4743594581, G4900274166, G5240500704, G5285405813, G5290307715, Advanced super-resolution fluorescence STED microscopy of the cellular interior., G5790500060, EXC 2051: Balance of the Microverse, G6715939638, G696823884, G7288767457, G7870782475, G80795443, Advanced Microscopy for Chromosome and RNA Dynamics., G8645460060, G8839205177, G984412895
OpenAlex ID: W4414622851