Abstract
Cartilage and bone are specialized skeletal tissues composed of unique extracellular matrices. Bone, in particular, has a highly calcified or mineralized matrix that makes microtomy and standard histological studies very challenging. Therefore, methods to appropriately fix and decalcify mineralized skeletal tissues have been developed to allow for paraffin processing and standard microtomy. In this chapter, we illustrate methods for tissue grossing, fixation, decalcification, paraffin processing, embedding, sectioning, and routine histological staining of demineralized murine skeletal tissues. We also discuss methods for decalcified frozen sectioning of skeletal tissues with and without the use of a tape-transfer system.
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References
Boskey AL (1992) Mineral-matrix interactions in bone and cartilage. Clin Orthop Relat Res 281:244–274
Erben RG (1997) Embedding of bone samples in methylmethacrylate: an improved method suitable for bone histomorphometry, histochemistry, and immunohistochemistry. J Histochem Cytochem 45(2):307–313
Merrell GA, Troiano NW, Coady CE, Kacena MA (2005) Effects of long-term fixation on histological quality of undecalcified murine bones embedded in methylmethacrylate. Biotech Histochem 80(3–4):139–146
Belluoccio D, Rowley L, Little CB, Bateman JF (2013) Maintaining mRNA integrity during decalcification of mineralized tissues. PLoS One 8(3):e58154
Shibata Y, Fujita S, Takahashi H, Yamaguchi A, Koji T (2000) Assessment of decalcifying protocols for detection of specific RNA by non-radioactive in situ hybridization in calcified tissues. Histochem Cell Biol 113(3):153–159
Bourque WT, Gross M, Hall BK (1993) A histological processing technique that preserves the integrity of calcified tissues (bone, enamel), yolky amphibian embryos, and growth factor antigens in skeletal tissue. J Histochem Cytochem 41(9):1429–1434
Callis GM (2008) “8/Bone.” The theory and practice of histological techniques, 6th edn. Churchill Livingston Elsevier, London, pp 333–348
Callis G, Sterchi D (2002) Animal processing manual. National Society for Histotechnology, Bowie, MD
Salie R, Li H, Jiang X, Rowe DW, Kalajzic I, Susa M (2008) A rapid, nonradioactive in situ hybridization technique for use on cryosectioned adult mouse bone. Calcif Tissue Int 83(3):212–221
Acknowledgments
We would like to thank Ryan Tierney for the initial development of some of these protocols. This work was supported in part by an NIH P30 Core Centers Grant (AR061307) and R01 grants (AR057022 and AR063071) to MJH.
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Mack, S.A., Maltby, K.M., Hilton, M.J. (2014). Demineralized Murine Skeletal Histology. In: Hilton, M. (eds) Skeletal Development and Repair. Methods in Molecular Biology, vol 1130. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-989-5_12
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DOI: https://doi.org/10.1007/978-1-62703-989-5_12
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-988-8
Online ISBN: 978-1-62703-989-5
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