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Intravital Imaging of Mouse Bone Marrow: Hemodynamics and Vascular Permeability

  • Yookyung Jung
  • Joel A. Spencer
  • Anthony P. Raphael
  • Juwell W. Wu
  • Clemens Alt
  • Judith R. Runnels
  • Briaira Geiger
  • Charles P. Lin
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1763)

Abstract

The bone marrow is a unique microenvironment where blood cells are produced and released into the circulation. At the top of the blood cell lineage are the hematopoietic stem cells (HSC), which are thought to reside in close association with the bone marrow vascular endothelial cells (Morrison and Scadden, Nature 505:327–334, 2014). Recent efforts at characterizing the HSC niche have prompted us to make close examinations of two distinct types of blood vessel in the bone marrow, the arteriolar vessels originating from arteries and sinusoidal vessels connected to veins. We found the two vessel types to exhibit different vascular permeabilites, hemodynamics, cell trafficking behaviors, and oxygen content (Itkin et al., Nature 532:323–328, 2016; Spencer et al., Nature 508:269–273, 2014). Here, we describe a method to quantitatively measure the permeability and hemodynamics of arterioles and sinusoids in murine calvarial bone marrow using intravital microscopy.

Key words

Bone marrow blood vessel Arterioles Sinusoids Permeability Hemodynamics Flow speed Blood vessel diameter Mouse restraint Intravital imaging 

Notes

Acknowledgement

This work is supported in part by NIH grant EB017274, DK103074, and HL 095489 (to C.P.L.), by IBS-R023-Y1 (to Y.J.), and by the Australian National Health and Medical Research Council (NHMRC), Early Career Fellowship #APP1088318 (to A.P.R.).

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Yookyung Jung
    • 1
    • 3
  • Joel A. Spencer
    • 1
    • 2
    • 4
  • Anthony P. Raphael
    • 1
    • 5
  • Juwell W. Wu
    • 1
  • Clemens Alt
    • 1
  • Judith R. Runnels
    • 1
  • Briaira Geiger
    • 6
  • Charles P. Lin
    • 1
  1. 1.Center for Systems Biology and Wellman Center for PhotomedicineMassachusetts General Hospital and Harvard Medical SchoolBostonUSA
  2. 2.Center for Regenerative MedicineMassachusetts General Hospital and Harvard Medical SchoolBostonUSA
  3. 3.Center for Molecular Spectroscopy and DynamicsInstitute for Basic Science (IBS)SeoulRepublic of Korea
  4. 4.School of EngineeringUniversity of California MercedMercedUSA
  5. 5.Dermatology Research CentreTranslational Research Institute, School of Medicine, The University of QueenslandSt LuciaAustralia
  6. 6.Department of ChemistryRichard Stockton College of New JerseyGallowayUSA

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