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Laser Capture Microdissection pp 191–202Cite as

Laser Capture Microdissection and Transcriptional Analysis of Sub-Populations of the Osteoblast Lineage from Undecalcified Bone

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1723))

Abstract

Transcriptional analysis of tissue samples is a useful and widely applied approach to provide insight into the molecular mechanisms engaged in response to phenotypic changes or external stimuli. We describe a method that overcomes the technical challenges associated with the application of Laser Capture Microdissection to undecalcified bone enabling us to collect high-quality bone tissue with maintained cellular morphology that is suitable for cryosectioning, fixation, and cutting. Using this method, we obtain samples enriched for specific cell types from the mature osteoblast lineage (osteoblasts, lining cells, i.e., quiescent osteoblasts, and osteocytes). RNA is well preserved and following extraction and amplification can be used as input to both low and high-throughput RNA analysis formats.

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Acknowledgments

The authors are grateful to Dr. Rogely Boyce of Amgen, Inc. for her careful review of this work.

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Authors and Affiliations

  1. Comparative Biology and Safety Sciences, Amgen Inc., Thousand Oaks, CA, USA

    Efrain Pacheco, Rong Hu & Scott Taylor

Authors
  1. Efrain Pacheco
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  2. Rong Hu
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  3. Scott Taylor
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Corresponding author

Correspondence to Scott Taylor .

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Editors and Affiliations

  1. School of Medicine, Medical Sciences and Nutrition, University of Aberdeen, Aberdeen, United Kingdom

    Graeme I. Murray

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Pacheco, E., Hu, R., Taylor, S. (2018). Laser Capture Microdissection and Transcriptional Analysis of Sub-Populations of the Osteoblast Lineage from Undecalcified Bone. In: Murray, G. (eds) Laser Capture Microdissection. Methods in Molecular Biology, vol 1723. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7558-7_10

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  • DOI: https://doi.org/10.1007/978-1-4939-7558-7_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7557-0

  • Online ISBN: 978-1-4939-7558-7

  • eBook Packages: Springer Protocols

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