Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope
Lateral organization of biological membranes is frequently studied using fluorescence microscopy. One of the most widely used probes for these studies is 2-dimethylamino- 6-lauroylnaphthalene (laurdan). The fluorescence of this probe is sensitive to the environment polarity, and thus laurdan reports...
Guardado en:
| Autores principales: | , , |
|---|---|
| Formato: | JOUR |
| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_00222275_v53_n3_p609_DodesTraian |
| Aporte de: |
| id |
todo:paper_00222275_v53_n3_p609_DodesTraian |
|---|---|
| record_format |
dspace |
| spelling |
todo:paper_00222275_v53_n3_p609_DodesTraian2023-10-03T14:28:31Z Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope Dodes Traian, M.M. González Flecha, F.L. Levi, V. Confocal microscopy Generalized polarization Lipid phases laurdan liposome article cell labeling cell vacuole confocal microscopy controlled study excitation fluorescence analysis fluorescence microscopy fluorescence polarization image processing lipid analysis lipid membrane lipid vesicle membrane fluidity nonhuman priority journal quantitative analysis 2-Naphthylamine Animals Cell Membrane Laurates Microscopy, Confocal Microscopy, Fluorescence Unilamellar Liposomes Lateral organization of biological membranes is frequently studied using fluorescence microscopy. One of the most widely used probes for these studies is 2-dimethylamino- 6-lauroylnaphthalene (laurdan). The fluorescence of this probe is sensitive to the environment polarity, and thus laurdan reports the local penetration of water when inserted in membranes. Unfortunately, this probe can only be used under two-photon excitation due to its low photostability. This is a very important limitation, because there are not too many laboratories with capability for two-photon microscopy. In this work, we explored the performance of 6-dodecanoyl-2-[N-methyl-N-(carboxymethyl)amino]naphthalene (C-laurdan), a carboxyl-modified version of laurdan, for imaging biological membranes using a conventional confocal micros copy setup. We acquired generalized polarization (GP) images of C-laurdan inserted in giant unillamelar vesicles composed of binary mixtures of lipids and verified that the probe allows observing the coexistence of different phases. We also tested the performance of the probe for measurement with living cells and registered GP images of melanophore cells labeled with C-laurdan in which we could observe highly ordered regions such as filopodia. These findings show that C-laurdan can be successfully employed for studies of membrane lateral organization using a conventional confocal microscope and can open the possibility of studying a wide variety of membrane-related processes. Copyright © 2012 by the American Society for Biochemistry and Molecular Biology, Inc. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00222275_v53_n3_p609_DodesTraian |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Confocal microscopy Generalized polarization Lipid phases laurdan liposome article cell labeling cell vacuole confocal microscopy controlled study excitation fluorescence analysis fluorescence microscopy fluorescence polarization image processing lipid analysis lipid membrane lipid vesicle membrane fluidity nonhuman priority journal quantitative analysis 2-Naphthylamine Animals Cell Membrane Laurates Microscopy, Confocal Microscopy, Fluorescence Unilamellar Liposomes |
| spellingShingle |
Confocal microscopy Generalized polarization Lipid phases laurdan liposome article cell labeling cell vacuole confocal microscopy controlled study excitation fluorescence analysis fluorescence microscopy fluorescence polarization image processing lipid analysis lipid membrane lipid vesicle membrane fluidity nonhuman priority journal quantitative analysis 2-Naphthylamine Animals Cell Membrane Laurates Microscopy, Confocal Microscopy, Fluorescence Unilamellar Liposomes Dodes Traian, M.M. González Flecha, F.L. Levi, V. Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope |
| topic_facet |
Confocal microscopy Generalized polarization Lipid phases laurdan liposome article cell labeling cell vacuole confocal microscopy controlled study excitation fluorescence analysis fluorescence microscopy fluorescence polarization image processing lipid analysis lipid membrane lipid vesicle membrane fluidity nonhuman priority journal quantitative analysis 2-Naphthylamine Animals Cell Membrane Laurates Microscopy, Confocal Microscopy, Fluorescence Unilamellar Liposomes |
| description |
Lateral organization of biological membranes is frequently studied using fluorescence microscopy. One of the most widely used probes for these studies is 2-dimethylamino- 6-lauroylnaphthalene (laurdan). The fluorescence of this probe is sensitive to the environment polarity, and thus laurdan reports the local penetration of water when inserted in membranes. Unfortunately, this probe can only be used under two-photon excitation due to its low photostability. This is a very important limitation, because there are not too many laboratories with capability for two-photon microscopy. In this work, we explored the performance of 6-dodecanoyl-2-[N-methyl-N-(carboxymethyl)amino]naphthalene (C-laurdan), a carboxyl-modified version of laurdan, for imaging biological membranes using a conventional confocal micros copy setup. We acquired generalized polarization (GP) images of C-laurdan inserted in giant unillamelar vesicles composed of binary mixtures of lipids and verified that the probe allows observing the coexistence of different phases. We also tested the performance of the probe for measurement with living cells and registered GP images of melanophore cells labeled with C-laurdan in which we could observe highly ordered regions such as filopodia. These findings show that C-laurdan can be successfully employed for studies of membrane lateral organization using a conventional confocal microscope and can open the possibility of studying a wide variety of membrane-related processes. Copyright © 2012 by the American Society for Biochemistry and Molecular Biology, Inc. |
| format |
JOUR |
| author |
Dodes Traian, M.M. González Flecha, F.L. Levi, V. |
| author_facet |
Dodes Traian, M.M. González Flecha, F.L. Levi, V. |
| author_sort |
Dodes Traian, M.M. |
| title |
Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope |
| title_short |
Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope |
| title_full |
Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope |
| title_fullStr |
Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope |
| title_full_unstemmed |
Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope |
| title_sort |
imaging lipid lateral organization in membranes with c-laurdan in a confocal microscope |
| url |
http://hdl.handle.net/20.500.12110/paper_00222275_v53_n3_p609_DodesTraian |
| work_keys_str_mv |
AT dodestraianmm imaginglipidlateralorganizationinmembraneswithclaurdaninaconfocalmicroscope AT gonzalezflechafl imaginglipidlateralorganizationinmembraneswithclaurdaninaconfocalmicroscope AT leviv imaginglipidlateralorganizationinmembraneswithclaurdaninaconfocalmicroscope |
| _version_ |
1807316124784656384 |