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Single-molecule microscopy reveals membrane microdomain organization of cells in a living vertebrate

Schaaf, Marcel J. M. ; Koopmans, Wiepke J. A. ; Meckel, Tobias ; Noort, John van ; Snaar-Jagalska, B. Ewa ; Schmidt, Thomas S. ; Spaink, Herman P. (2009)
Single-molecule microscopy reveals membrane microdomain organization of cells in a living vertebrate.
In: Biophysical journal, 97 (4)
Artikel, Bibliographie

Kurzbeschreibung (Abstract)

It has been possible for several years to study the dynamics of fluorescently labeled proteins by single-molecule microscopy, but until now this technology has been applied only to individual cells in culture. In this study, it was extended to stem cells and living vertebrate organisms. As a molecule of interest we used yellow fluorescent protein fused to the human H-Ras membrane anchor, which has been shown to serve as a model for proteins anchored in the plasma membrane. We used a wide-field fluorescence microscopy setup to visualize individual molecules in a zebrafish cell line (ZF4) and in primary embryonic stem cells. A total-internal-reflection microscopy setup was used for imaging in living organisms, in particular in epidermal cells in the skin of 2-day-old zebrafish embryos. Our results demonstrate the occurrence of membrane microdomains in which the diffusion of membrane proteins in a living organism is confined. This membrane organization differed significantly from that observed in cultured cells, illustrating the relevance of performing single-molecule microscopy in living organisms.

Typ des Eintrags: Artikel
Erschienen: 2009
Autor(en): Schaaf, Marcel J. M. ; Koopmans, Wiepke J. A. ; Meckel, Tobias ; Noort, John van ; Snaar-Jagalska, B. Ewa ; Schmidt, Thomas S. ; Spaink, Herman P.
Art des Eintrags: Bibliographie
Titel: Single-molecule microscopy reveals membrane microdomain organization of cells in a living vertebrate
Sprache: Englisch
Publikationsjahr: 2009
Titel der Zeitschrift, Zeitung oder Schriftenreihe: Biophysical journal
Jahrgang/Volume einer Zeitschrift: 97
(Heft-)Nummer: 4
Kurzbeschreibung (Abstract):

It has been possible for several years to study the dynamics of fluorescently labeled proteins by single-molecule microscopy, but until now this technology has been applied only to individual cells in culture. In this study, it was extended to stem cells and living vertebrate organisms. As a molecule of interest we used yellow fluorescent protein fused to the human H-Ras membrane anchor, which has been shown to serve as a model for proteins anchored in the plasma membrane. We used a wide-field fluorescence microscopy setup to visualize individual molecules in a zebrafish cell line (ZF4) and in primary embryonic stem cells. A total-internal-reflection microscopy setup was used for imaging in living organisms, in particular in epidermal cells in the skin of 2-day-old zebrafish embryos. Our results demonstrate the occurrence of membrane microdomains in which the diffusion of membrane proteins in a living organism is confined. This membrane organization differed significantly from that observed in cultured cells, illustrating the relevance of performing single-molecule microscopy in living organisms.

Fachbereich(e)/-gebiet(e): 10 Fachbereich Biologie
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10 Fachbereich Biologie > Plant Membrane Biophyscis (am 20.12.23 umbenannt in Biologie der Algen und Protozoen)
10 Fachbereich Biologie > Membrane Dynamics
Hinterlegungsdatum: 06 Jun 2011 13:30
Letzte Änderung: 26 Jul 2021 07:31
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