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Fluorescence Correlation and Cross-Correlation Spectroscopy in Zebrafish

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Morphogen Gradients

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1863))

Abstract

There has been increasing interest in biophysical studies on live organisms to gain better insights into physiologically relevant biological events at the molecular level. Zebrafish (Danio rerio) is a viable vertebrate model to study such events due to its genetic and evolutionary similarities to humans, amenability to less invasive fluorescence techniques owing to its transparency and well-characterized genetic manipulation techniques. Fluorescence techniques used to probe biomolecular dynamics and interactions of molecules in live zebrafish embryos are therefore highly sought-after to bridge molecular and developmental events. Fluorescence correlation and cross-correlation spectroscopy (FCS and FCCS) are two robust techniques that provide molecular level information on dynamics and interactions respectively. Here, we detail the steps for applying confocal FCS and FCCS, in particular single-wavelength FCCS (SW-FCCS), in live zebrafish embryos, beginning with sample preparation, instrumentation, calibration, and measurements on the FCS/FCCS instrument and ending with data analysis.

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Change history

  • 07 December 2018

    The book was published with the following errors: In Chapter 1, figure 1 was stretched. This misconfiguration has now been modified.

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Acknowledgment

X.W.N. is supported by the NUS graduate research scholarship. T.W. acknowledges funding by the Ministry of Education of Singapore (grant number MOE2016-T3-1-005). Work in the laboratory of K.S. is supported by Warwick Medical School and the BBSRC. K.S. thanks Andreas Zaucker and Scott Clarke for the image of the experimental setup for mounting zebrafish embryos.

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Authors and Affiliations

  1. Department of Chemistry and Centre for Bioimaging Sciences, National University of Singapore, Singapore, Singapore

    Xue Wen Ng & Thorsten Wohland

  2. Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry, UK

    Karuna Sampath

  3. Department of Biological Sciences, National University of Singapore, Singapore, Singapore

    Thorsten Wohland

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  1. Xue Wen Ng
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  2. Karuna Sampath
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  3. Thorsten Wohland
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Correspondence to Thorsten Wohland .

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Editors and Affiliations

  1. Integrated Microscopy Core, Baylor College of Medicine, Houston, TX, USA

    Julien Dubrulle

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Ng, X.W., Sampath, K., Wohland, T. (2018). Fluorescence Correlation and Cross-Correlation Spectroscopy in Zebrafish. In: Dubrulle, J. (eds) Morphogen Gradients. Methods in Molecular Biology, vol 1863. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8772-6_5

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  • DOI: https://doi.org/10.1007/978-1-4939-8772-6_5

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