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Abstract:

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.

Registro:

Documento: Artículo
Título:Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope
Autor:Dodes Traian, M.M.; González Flecha, F.L.; Levi, V.
Filiación:Departamento de Química Biológica, Universidad de Buenos Aires, Ciudad Universitaria, CP 1428 Ciudad de Buenos Aires, Argentina
Laboratorio de Biofísica Molecular, Universidad de Buenos Aires, CONICET, Buenos Aires, Argentina
Palabras clave: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
Año:2012
Volumen:53
Número:3
Página de inicio:609
Página de fin:616
DOI: http://dx.doi.org/10.1194/jlr.D021311
Título revista:Journal of Lipid Research
Título revista abreviado:J. Lipid Res.
ISSN:00222275
CODEN:JLPRA
CAS:laurdan, 74515-25-6; 2-Naphthylamine, 91-59-8; Laurates; Unilamellar Liposomes; laurdan, 74515-25-6
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00222275_v53_n3_p609_DodesTraian

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Citas:

---------- APA ----------
Dodes Traian, M.M., González Flecha, F.L. & Levi, V. (2012) . Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope. Journal of Lipid Research, 53(3), 609-616.
http://dx.doi.org/10.1194/jlr.D021311
---------- CHICAGO ----------
Dodes Traian, M.M., González Flecha, F.L., Levi, V. "Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope" . Journal of Lipid Research 53, no. 3 (2012) : 609-616.
http://dx.doi.org/10.1194/jlr.D021311
---------- MLA ----------
Dodes Traian, M.M., González Flecha, F.L., Levi, V. "Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope" . Journal of Lipid Research, vol. 53, no. 3, 2012, pp. 609-616.
http://dx.doi.org/10.1194/jlr.D021311
---------- VANCOUVER ----------
Dodes Traian, M.M., González Flecha, F.L., Levi, V. Imaging lipid lateral organization in membranes with C-laurdan in a confocal microscope. J. Lipid Res. 2012;53(3):609-616.
http://dx.doi.org/10.1194/jlr.D021311