ZFIN ID: ZDB-PUB-091120-13
Probing events with single molecule sensitivity in zebrafish and Drosophila embryos by fluorescence correlation spectroscopy
Shi, X., Shin Teo, L., Pan, X., Chong, S.W., Kraut, R., Korzh, V., and Wohland, T.
Date: 2009
Source: Developmental dynamics : an official publication of the American Association of Anatomists   238(12): 3156-3167 (Journal)
Registered Authors: Chong, Shang Wei, Korzh, Vladimir
Keywords: fluorescence correlation spectroscopy, zebrafish, Drosophila, diffusion coefficient, blood flow velocity, autofluorescence, penetration depth
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Diffusion
  • Drosophila melanogaster/embryology*
  • Drosophila melanogaster/genetics
  • Drosophila melanogaster/metabolism
  • Embryo, Nonmammalian/blood supply
  • Embryonic Development/physiology*
  • Female
  • Fluorescence
  • Fluorescent Antibody Technique/instrumentation
  • Fluorescent Antibody Technique/methods
  • Green Fluorescent Proteins/genetics
  • Green Fluorescent Proteins/metabolism
  • Male
  • Models, Biological
  • Regional Blood Flow/physiology
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence/methods
  • Zebrafish/embryology*
  • Zebrafish/genetics
  • Zebrafish/metabolism
PubMed: 19882725 Full text @ Dev. Dyn.
FIGURES
ABSTRACT
Zebrafish and Drosophila are animal models widely used in developmental biology. High-resolution microscopy and live imaging techniques have allowed the investigation of biological processes down to the cellular level in these models. Here, using fluorescence correlation spectroscopy (FCS), we show that even processes on a molecular level can be studied in these embryos. The two animal models provide different advantages and challenges. We first characterize their autofluorescence pattern and determine usable penetration depth for FCS especially in the case of zebrafish, where tissue thickness is an issue. Next, the applicability of FCS to study molecular processes is shown by the determination of blood flow velocities with high spatial resolution and the determination of diffusion coefficients of cytosolic and membrane-bound enhanced green fluorescent protein-labeled proteins in different cell types. This work provides an approach to study molecular processes in vivo and opens up the possibility to relate these molecular processes to developmental biology questions.
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