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Hematopoietic Stem Cell Protocols pp 247–265Cite as

Intravital Imaging of Hematopoietic Stem Cells in the Mouse Skull

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1185))

Abstract

Over the past 50 years, much insight has been gained into the biology of hematopoietic stem cells (HSCs). Much of this information has been gained though isolation of specific bone marrow populations, and transplantation into irradiated recipients followed by characterization of chimeras months later. These studies have yielded insights into the function of HSCs, but have shed little light on the interactions of individual stem cells with their environment. Characterization of the behavior of single HSCs awaited the use of relatively noninvasive intravital microscopy, which allows one to identify rare cells in real time and follow them in multiple imaging sessions. Here we describe techniques used to image transplanted HSCs in the mouse calvarium using hybrid confocal/multi-photon microscopy and second harmonic imaging. For detection, fluorescently tagged HSCs are transplanted into a recipient mouse. The architecture of the bone marrow can be delineated using a combination of fluorescent probes and vascular dyes, second harmonic generation to detect the collagen signal from bone, and transgenic recipient mice containing specific fluorescent support cell populations.

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Acknowledgments

The authors wish to thank Raphaël Turcotte for the mouse skull shown in Fig. 2a and Drs. Luke Mortensen and Amir Schajnovitz for the image displayed in Fig. 2b. This work was funded by NIH HL097748, HL97794, and HL100402.

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Authors and Affiliations

  1. Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    Juwell W. Wu, Judith M. Runnels & Charles P. Lin

  2. Wellman Center for Photomedicine, Massachusetts General Hospital, 185 Cambridge Street, CPZN8238, Boston, MA, 02114, USA

    Juwell W. Wu, Judith M. Runnels & Charles P. Lin

Authors
  1. Juwell W. Wu
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  2. Judith M. Runnels
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  3. Charles P. Lin
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Correspondence to Charles P. Lin .

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Editors and Affiliations

  1. Department of Pediatrics, Aflac Cancer and Blood Disorders Center Emory University School of Medicine, Atlanta, Georgia, USA

    Kevin D. Bunting

  2. Department of Pediatrics, Emory University School of Medicine Aflac Cancer & Blood Disorders Ctr, Atlanta, Georgia, USA

    Cheng-Kui Qu

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Wu, J.W., Runnels, J.M., Lin, C.P. (2014). Intravital Imaging of Hematopoietic Stem Cells in the Mouse Skull. In: Bunting, K., Qu, CK. (eds) Hematopoietic Stem Cell Protocols. Methods in Molecular Biology, vol 1185. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1133-2_17

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  • DOI: https://doi.org/10.1007/978-1-4939-1133-2_17

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1132-5

  • Online ISBN: 978-1-4939-1133-2

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