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Cell Signaling Reactions pp 183–197Cite as

In Vivo Single-Molecule Microscopy Using the Zebrafish Model System

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Abstract

In recent years, several groups have succeeded in extending single-molecule microscopy technology to the level of a living vertebrate organism using the zebrafish embryo as a model system. In this chapter an overview will be presented of these studies and three lines of research will be discussed. First, work will be presented in which fluorescent proteins have been imaged at the single-molecule level in the epidermis of zebrafish embryos using total internal reflection fluorescence (TIRF) microscopy. Second, investigations will be presented in which individual quantum dots have been imaged in zebrafish embryos by selective plane illumination microscopy (SPIM). Third, studies will be discussed in which fluorescence correlation spectroscopy (FCS) has been applied to fluorescent proteins in zebrafish embryos. All three research lines show discrepancies between results obtained in zebrafish embryos and data obtained in cell cultures, illustrating the relevance of performing these studies in an in vivo model.

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Abbreviations

CCD:

charge-coupled device

CHO:

Chinese hamster ovary

dpf:

days post fertilization

eYFP:

enhanced yellow fluorescent protein

FCS:

fluorescence correlation spectroscopy

Fgf:

fibroblast growth factor

GFP:

green fluorescent protein

hpf:

hours post fertilization

HSPG:

heparan sulphate proteoglycan

IQGAP1:

IQ motif containing GTPase activating protein 1

LCK:

lymphocyte-specific protein tyrosine kinase

mRFP:

monomeric red fluorescent protein

PTU:

phenylthiourea

QD:

quantum dot

SPIM:

selective plane illumination microscopy

SW-FCCS:

Single Wavelength Fluorescence Cross-Correlation Spectroscopy

TIRF:

total internal reflection fluorescence

TMR:

tetramethylrhodamine

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Author information

Authors and Affiliations

  1. Molecular Cell Biology, Institute of Biology, Leiden University, Einsteinweg 55, 2333CC, Leiden, The Netherlands

    Marcel J. M. Schaaf

  2. Physics of Life Processes, Institute of Physics, Leiden University, Niels Bohrweg 2, 2333CA, Leiden, The Netherlands

    Thomas S. Schmidt

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  1. Marcel J. M. Schaaf
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Correspondence to Marcel J. M. Schaaf .

Editor information

Editors and Affiliations

  1. , Cellular Informatics Laboratory, RIKEN, Hirosawa 2-1, Wako, 351-0198, Japan

    Yasushi Sako

  2. Graduate School of Frontier Biosciences, Laboratories of Nanobiology, Osaka University, Yamadaoka 1-3, Suita, 565-871, Japan

    Masahiro Ueda

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Schaaf, M.J.M., Schmidt, T.S. (2011). In Vivo Single-Molecule Microscopy Using the Zebrafish Model System. In: Sako, Y., Ueda, M. (eds) Cell Signaling Reactions. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9864-1_9

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