title: Probing events with single molecule sensitivity in zebrafish and Drosophila embryos by fluorescence correlation spectroscopy.
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.
authors.affiliation: Department of Chemistry, National University of Singapore, Singapore.
authors.forename: Xianke
authors.initials: X
authors.lastname: Shi
authors.forename: Lin Shin
authors.initials: LS
authors.lastname: Teo
authors.forename: Xiaotao
authors.initials: X
authors.lastname: Pan
authors.forename: Shang-Wei
authors.initials: SW
authors.lastname: Chong
authors.forename: Rachel
authors.initials: R
authors.lastname: Kraut
authors.forename: Vladimir
authors.initials: V
authors.lastname: Korzh
authors.forename: Thorsten
authors.initials: T
authors.lastname: Wohland
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004058:Diffusion; D004331:Drosophila melanogaster / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000098:blood supply; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D005453:Fluorescence; D005455:Fluorescent Antibody Technique / Q000295:instrumentation / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008954:Models, Biological; D012039:Regional Blood Flow / Q000502:physiology; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics / Q000378:metabolism
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins