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Measuring bone blood supply in mice using fluorescent microspheres

Nature Protocols volume 4, pages 1749–1758 (2009)Cite this article

Abstract

Fluorescent microspheres are commonly used to assess bone blood supply in large animals, but the technique is not widely used in smaller mammals, as traditional methods such as reference blood sampling, ventilation and catheterization are not easily applied. This protocol describes a viable alternative for measuring bone and organ perfusion in mice using modified fluorescent microsphere techniques. Microspheres are injected directly into the left heart and a reference tissue is used to calculate relative bone and organ blood supply. On the basis of a sample of 15 mice with 5 tissues each, the entire protocol takes 140.5 h to complete from animal preparation through statistical analysis. This timing includes 72 h of mandated pauses for bone decalcification and digestion, as well as 48 h for data analysis. Exclusive of pauses or additional analyses that could increase the time required, this protocol takes 20.5 h bench time to complete.

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Figure 1: Relative microsphere deposition in the tibia.
Figure 2: Flowchart illustrating the fluorescent microsphere injection and recovery procedures used here.

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Acknowledgements

This research was supported by a grant from the National Science Foundation (BCS-0524899). W. Horne, J. Jacobi, D. Rurak, R. Glenny and the Fluorescent Microsphere Resource Center provided valuable input on microsphere injection and recovery techniques, and A. Carey (Sefar America) helped obtain essential filtration material. Special thanks to O. Lovejoy, D. King, W.E. Horton, C. Vinyard, J. Stalvey, R. Meindl, P. Reno, C. Farnum and two anonymous reviewers for critical suggestions and to D. McBurney, B. Armfield, E. Bailey, N. Smallwood and J. Hardwick for advice and technical assistance in developing this protocol.

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Authors and Affiliations

  1. Department of Biomedical Sciences, Kent State University, Kent Ohio, USA.,

    Maria A Serrat

  2. Department of Anatomy, Northeastern Ohio Universities College of Medicine, Rootstown, Ohio, USA

    Maria A Serrat

  3. Department of Anatomy and Pathology, Joan C. Edwards School of Medicine, Marshall University, Huntington, West Virginia, USA

    Maria A Serrat

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  1. Maria A Serrat
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Correspondence to Maria A Serrat.

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Serrat, M. Measuring bone blood supply in mice using fluorescent microspheres. Nat Protoc 4, 1749–1758 (2009). https://doi.org/10.1038/nprot.2009.190

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