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
-
- Embryonic Development/physiology*
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/metabolism
- Male
- Sensitivity and Specificity
- Female
- Fluorescent Antibody Technique/instrumentation
- Fluorescent Antibody Technique/methods
- Regional Blood Flow/physiology
- Fluorescence
- Spectrometry, Fluorescence/methods
- Embryo, Nonmammalian/blood supply
- Zebrafish/embryology*
- Zebrafish/genetics
- Zebrafish/metabolism
- Animals, Genetically Modified
- Diffusion
- Models, Biological
- Animals
- Drosophila melanogaster/embryology*
- Drosophila melanogaster/genetics
- Drosophila melanogaster/metabolism
- PubMed
- 19882725 Full text @ Dev. Dyn.
Citation
Shi, X., Shin Teo, L., Pan, X., Chong, S.W., Kraut, R., Korzh, V., and Wohland, T. (2009) Probing events with single molecule sensitivity in zebrafish and Drosophila embryos by fluorescence correlation spectroscopy. Developmental Dynamics : an official publication of the American Association of Anatomists. 238(12):3156-3167.
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
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping