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Ex Utero Culture and Imaging of Mouse Embryos

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Vertebrate Embryogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1920))

Abstract

Mouse genetic approaches when combined with live imaging tools are revolutionizing our current understanding of mammalian developmental biology. The availability and improvement of a wide variety of genetically encoded fluorescent proteins have provided indispensable tools to visualize cells and subcellular features in living organisms. It is now possible to generate genetically modified mouse lines expressing several spectrally distinct fluorescent proteins in a tissue-specific or -inducible manner. Such reporter-expressing lines make it possible to image dynamic cellular behaviors in the context of living embryos undergoing normal or aberrant development. As with all viviparous mammals, mouse embryos develop within the uterus, and so live imaging experiments require culture conditions that closely mimic the in vivo environment. Over the past decades, significant advances have been made in developing conditions for culturing both pre- and postimplantation-stage mouse embryos. In this chapter, we discuss routine methods for ex utero culture of preimplantation- and postimplantation-stage mouse embryos. In particular, we describe protocols for collecting mouse embryos of various stages, setting up culture conditions for their ex utero culture and imaging, and using laser scanning confocal microscopy to visualize live processes in mouse embryos expressing fluorescent reporters.

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Acknowledgments

We thank our present and past laboratory colleagues for helping perfect the techniques detailed in this chapter. Work in our laboratory is supported by the National Institutes of Health (R01DK084391, R01HD094868, and P30CA008748) and The Starr Foundation.

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Authors and Affiliations

  1. Developmental Biology Program, Sloan Kettering Institute, Memorial Sloan Kettering Cancer Center, New York, NY, USA

    Sonja Nowotschin, Vidur Garg & Anna-Katerina Hadjantonakis

  2. Department of Experimental Embryology, Institute of Genetics and Animal Breeding, Polish Academy of Sciences, Jastrzębiec, Poland

    Anna Piliszek

Authors
  1. Sonja Nowotschin
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  2. Vidur Garg
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  3. Anna Piliszek
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  4. Anna-Katerina Hadjantonakis
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Corresponding author

Correspondence to Anna-Katerina Hadjantonakis .

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Editors and Affiliations

  1. Laboratory of Genetics, University of Wisconsin–Madison, Madison, WI, USA

    Francisco J. Pelegri

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Nowotschin, S., Garg, V., Piliszek, A., Hadjantonakis, AK. (2019). Ex Utero Culture and Imaging of Mouse Embryos. In: Pelegri, F. (eds) Vertebrate Embryogenesis. Methods in Molecular Biology, vol 1920. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9009-2_11

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  • DOI: https://doi.org/10.1007/978-1-4939-9009-2_11

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