RRC ID 6236
Author Shi X, Teo LS, Pan X, Chong SW, Kraut R, Korzh V, Wohland T.
Title Probing events with single molecule sensitivity in zebrafish and Drosophila embryos by fluorescence correlation spectroscopy.
Journal 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.
Volume 238(12)
Pages 3156-67
Published 2009-12-1
DOI 10.1002/dvdy.22140
PMID 19882725
MeSH 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
IF 3.275
Times Cited 25
WOS Category ANATOMY & MORPHOLOGY DEVELOPMENTAL BIOLOGY
Resource
Zebrafish rw0(Tg(CM-isl1:GFP))