ZFIN ID: ZDB-PUB-090819-16
Single-molecule microscopy reveals membrane microdomain organization of cells in a living vertebrate
Schaaf, M.J., Koopmans, W.J., Meckel, T., van Noort, J., Snaar-Jagalska, B.E., Schmidt, T.S., and Spaink, H.P.
Date: 2009
Source: Biophysical journal   97(4): 1206-1214 (Journal)
Registered Authors: Schaaf, Marcel J. M., Snaar-Jagalska, Ewa B., Spaink, Herman P.
Keywords: none
MeSH Terms:
  • Animals
  • Cells, Cultured
  • Embryonic Stem Cells/cytology
  • Embryonic Stem Cells/ultrastructure*
  • Membrane Microdomains/ultrastructure*
  • Microscopy, Confocal/methods*
  • Microscopy, Fluorescence/methods*
  • Molecular Probe Techniques
  • Zebrafish/anatomy & histology*
  • Zebrafish/embryology*
PubMed: 19686669 Full text @ Biophys. J.
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.
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