PUBLICATION

Probing events with single molecule sensitivity in zebrafish and Drosophila embryos by fluorescence correlation spectroscopy

Authors
Shi, X., Shin Teo, L., Pan, X., Chong, S.W., Kraut, R., Korzh, V., and Wohland, T.
ID
ZDB-PUB-091120-13
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.
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.
Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes