Skip to main content
Book cover

Intravital Imaging of Dynamic Bone and Immune Systems pp 11–22Cite as

Intravital Imaging of Mouse Bone Marrow: Hemodynamics and Vascular Permeability

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

This is a preview of subscription content, access via your institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • DOI: 10.1007/978-1-4939-7762-8_2
  • Chapter length: 12 pages
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
eBook
USD   89.00
Price excludes VAT (USA)
  • ISBN: 978-1-4939-7762-8
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
Softcover Book
USD   109.99
Price excludes VAT (USA)
Hardcover Book
USD   149.99
Price excludes VAT (USA)
Learn about institutional subscriptions
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Spencer JA, Ferraro F, Roussakis E et al (2014) Direct measurement of local oxygen concentration in the bone marrow of live animals. Nature 508:269–273

    CrossRef  CAS  PubMed  PubMed Central  Google Scholar 

  2. Kunisaki Y et al (2013) Arteriolar niches maintain haematopoietic stem cell quiescence. Nature 502:637–643

    CrossRef  CAS  PubMed  PubMed Central  Google Scholar 

  3. Nombela-Arrieta C et al (2013) Quantitative imaging of haematopoietic stem and progenitor cell localization and hypoxic status in the bone marrow microenvironment. Nat Cell Biol 15:533–543

    CrossRef  CAS  PubMed  PubMed Central  Google Scholar 

  4. Li XM, Hu Z, Jorgenson ML, Slayton WB (2009) High levels of acetylated low-density lipoprotein uptake and low tyrosine kinase with immunoglobulin and epidermal growth factor homology domains-2 (Tie2) promoter activity distinguish sinusoids from other vessel types in murine bone marrow. Circulation 120:1910–1918

    CrossRef  CAS  PubMed  Google Scholar 

  5. Itkin T, Gur-Cohen S, Spencer JA et al (2016) Distinct bone marrow blood vessels differentially regulate haematopoiesis. Nature 532:323–328

    CrossRef  CAS  PubMed  Google Scholar 

  6. Veilleux I, Spencer JA, Biss DP et al (2008) In vivo cell tracking with video rate multimodality laser scanning microscopy. IEEE J Sel Top Quantum Electron 14:10–18

    CrossRef  CAS  Google Scholar 

  7. Lo Celso C, Lin CP, Scadden DT (2011) In vivo imaging of transplanted hematopoietic stem and progenitor cells in mouse calvarium bone marrow. Nat Protoc 6:1–14

    CrossRef  PubMed  Google Scholar 

  8. Wu JW, Runnels JM, Lin CP (2014) Intravital imaging of hematopoietic stem cells in the mouse skull. Methods Mol Biol 1185:247–265

    CrossRef  PubMed  Google Scholar 

  9. Morrison SJ, Scadden DT (2014) The bone marrow niche for haematopoietic stem cells. Nature 505:327–334

    CrossRef  CAS  PubMed  PubMed Central  Google Scholar 

  10. Schindelin J, Arganda-Carreras I, Frise E et al (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682

    CrossRef  CAS  PubMed  Google Scholar 

  11. Tseng Q, Wang I, Duchemin-Pelletier E et al (2011) A new micropatterning method of soft substrates reveals that different tumorigenic signals can promote or reduce cell contraction levels. Lab Chip 11:2231

    CrossRef  CAS  PubMed  Google Scholar 

  12. Schneider CA, Rasband WS, Eliceiri KW (2012) NIH image to ImageJ: 25 years of image analysis. Nat Methods 9:671–675

    CrossRef  CAS  PubMed  PubMed Central  Google Scholar 

  13. Berg HC (1993) Random walks in biology. Princeton University Press, Princeton

    Google Scholar 

Download references

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.).

Author information

Authors and Affiliations

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

    Yookyung Jung, Joel A. Spencer, Anthony P. Raphael, Juwell W. Wu, Clemens Alt, Judith R. Runnels & Charles P. Lin

  2. Center for Regenerative Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA

    Joel A. Spencer

  3. Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science (IBS), Seoul, Republic of Korea

    Yookyung Jung

  4. School of Engineering, University of California Merced, Merced, CA, 95343, USA

    Joel A. Spencer

  5. Dermatology Research Centre, Translational Research Institute, School of Medicine, The University of Queensland, St Lucia, Australia

    Anthony P. Raphael

  6. Department of Chemistry, Richard Stockton College of New Jersey, Galloway, NJ, USA

    Briaira Geiger

Authors
  1. Yookyung Jung
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Joel A. Spencer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anthony P. Raphael
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Juwell W. Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Clemens Alt
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Judith R. Runnels
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Briaira Geiger
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Charles P. Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Charles P. Lin .

Editor information

Editors and Affiliations

  1. Department of Immunology and Cell Biology, Graduate School of Medicine, Osaka University, Osaka, Osaka, Japan

    Masaru Ishii

Rights and permissions

Reprints and Permissions

Copyright information

© 2018 Springer Science+Business Media, LLC

About this protocol

Verify currency and authenticity via CrossMark

Cite this protocol

Jung, Y. et al. (2018). Intravital Imaging of Mouse Bone Marrow: Hemodynamics and Vascular Permeability. In: Ishii, M. (eds) Intravital Imaging of Dynamic Bone and Immune Systems . Methods in Molecular Biology, vol 1763. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7762-8_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7762-8_2

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7761-1

  • Online ISBN: 978-1-4939-7762-8

  • eBook Packages: Springer Protocols