Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes
Understanding the plasma membrane nanoscale organization and dynamics in living cells requires microscopy techniques with high spatial and temporal resolution that permit for long acquisition times and allow for the quantification of membrane biophysical properties, such as lipid ordering. Among the...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-12-01
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| Series: | Biophysical Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2667074721000239 |
| _version_ | 1818490810777206784 |
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| author | Pablo Carravilla Anindita Dasgupta Gaukhar Zhurgenbayeva Dmytro I. Danylchuk Andrey S. Klymchenko Erdinc Sezgin Christian Eggeling |
| author_facet | Pablo Carravilla Anindita Dasgupta Gaukhar Zhurgenbayeva Dmytro I. Danylchuk Andrey S. Klymchenko Erdinc Sezgin Christian Eggeling |
| author_sort | Pablo Carravilla |
| collection | DOAJ |
| description | Understanding the plasma membrane nanoscale organization and dynamics in living cells requires microscopy techniques with high spatial and temporal resolution that permit for long acquisition times and allow for the quantification of membrane biophysical properties, such as lipid ordering. Among the most popular super-resolution techniques, stimulated emission depletion (STED) microscopy offers one of the highest temporal resolutions, ultimately defined by the scanning speed. However, monitoring live processes using STED microscopy is significantly limited by photobleaching, which recently has been circumvented by exchangeable membrane dyes that only temporarily reside in the membrane. Here, we show that NR4A, a polarity-sensitive exchangeable plasma membrane probe based on Nile red, permits the super-resolved quantification of membrane biophysical parameters in real time with high temporal and spatial resolution as well as long acquisition times. The potential of this polarity-sensitive exchangeable dye is showcased by live-cell real-time three-dimensional STED recordings of bleb formation and lipid exchange during membrane fusion as well as by STED-fluorescence correlation spectroscopy experiments for the simultaneous quantification of membrane dynamics and lipid packing that correlate in model and live-cell membranes. |
| first_indexed | 2024-12-10T17:22:18Z |
| format | Article |
| id | doaj.art-1c5f5b996dbb433692525b93b8b6a560 |
| institution | Directory Open Access Journal |
| issn | 2667-0747 |
| language | English |
| last_indexed | 2024-12-10T17:22:18Z |
| publishDate | 2021-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biophysical Reports |
| spelling | doaj.art-1c5f5b996dbb433692525b93b8b6a5602022-12-22T01:39:56ZengElsevierBiophysical Reports2667-07472021-12-0112100023Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyesPablo Carravilla0Anindita Dasgupta1Gaukhar Zhurgenbayeva2Dmytro I. Danylchuk3Andrey S. Klymchenko4Erdinc Sezgin5Christian Eggeling6Leibniz Institute of Photonic Technology e.V., Jena, Germany; Faculty of Physics and Astronomy, Institute of Applied Optics and Biophysics, Friedrich Schiller University Jena, Jena, Germany; Corresponding authorLeibniz Institute of Photonic Technology e.V., Jena, Germany; Faculty of Physics and Astronomy, Institute of Applied Optics and Biophysics, Friedrich Schiller University Jena, Jena, GermanyFaculty of Physics and Astronomy, Institute of Applied Optics and Biophysics, Friedrich Schiller University Jena, Jena, Germany; Jena School for Microbial Communication, Friedrich Schiller University Jena, Jena, GermanyLaboratoire de Bioimagerie et Pathologies, UMR 7021 CNRS, Université de Strasbourg, Illkirch, FranceLaboratoire de Bioimagerie et Pathologies, UMR 7021 CNRS, Université de Strasbourg, Illkirch, FranceScience for Life Laboratory, Department of Women’s and Children’s Health, Karolinska Institutet, Stockholm, Sweden; Corresponding authorLeibniz Institute of Photonic Technology e.V., Jena, Germany; Faculty of Physics and Astronomy, Institute of Applied Optics and Biophysics, Friedrich Schiller University Jena, Jena, Germany; Jena School for Microbial Communication, Friedrich Schiller University Jena, Jena, Germany; Medical Research Council Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United Kingdom; Jena Center for Soft Matter, Jena, GermanyUnderstanding the plasma membrane nanoscale organization and dynamics in living cells requires microscopy techniques with high spatial and temporal resolution that permit for long acquisition times and allow for the quantification of membrane biophysical properties, such as lipid ordering. Among the most popular super-resolution techniques, stimulated emission depletion (STED) microscopy offers one of the highest temporal resolutions, ultimately defined by the scanning speed. However, monitoring live processes using STED microscopy is significantly limited by photobleaching, which recently has been circumvented by exchangeable membrane dyes that only temporarily reside in the membrane. Here, we show that NR4A, a polarity-sensitive exchangeable plasma membrane probe based on Nile red, permits the super-resolved quantification of membrane biophysical parameters in real time with high temporal and spatial resolution as well as long acquisition times. The potential of this polarity-sensitive exchangeable dye is showcased by live-cell real-time three-dimensional STED recordings of bleb formation and lipid exchange during membrane fusion as well as by STED-fluorescence correlation spectroscopy experiments for the simultaneous quantification of membrane dynamics and lipid packing that correlate in model and live-cell membranes.http://www.sciencedirect.com/science/article/pii/S2667074721000239 |
| spellingShingle | Pablo Carravilla Anindita Dasgupta Gaukhar Zhurgenbayeva Dmytro I. Danylchuk Andrey S. Klymchenko Erdinc Sezgin Christian Eggeling Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes Biophysical Reports |
| title | Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes |
| title_full | Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes |
| title_fullStr | Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes |
| title_full_unstemmed | Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes |
| title_short | Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes |
| title_sort | long term sted imaging of membrane packing and dynamics by exchangeable polarity sensitive dyes |
| url | http://www.sciencedirect.com/science/article/pii/S2667074721000239 |
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