Abstract
Laser capture microdissection (LCM) enables the removal of discrete microstructures or cell types from properly prepared histological sections. Extraction of RNA from microdissected tissue followed by quantitative reverse transcriptase-polymerase chain (QRT-PCR) reaction permits the analysis of cell-type or microstructure-specific gene expression changes that occur in response to various stimuli in the environment. In our lab, the combination of LCM and QRT-PCR has proven very useful in the determination of the in vivo gene expression changes that occur in bronchiolar epithelium in response to inhalation of crocidolite asbestos. A detailed description of the preparation of cDNA from bronchiolar epithelial cells obtained by LCM is described in this work.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Emmert-Buck, M. R., Bonner, R. F., Smith, P. D., et al. (1996) Laser capture microdissection. Science 274, 998–1001.
Taatjes, D. J., Palmer, C. J., Pantano, C., Hoffmann, S.B., Cummins, A., and Mossman, B.T. (2001) Laser-based microscopic approaches: application to cell signaling in environmental lung disease. Biotechniques 31, 880–894.
Manning, C. B., Cummins, A. B., Jung, M. W., et al. (2002) A mutant epidermal growth factor receptor targeted to lung epithelium inhibits asbestos-induced proliferation and proto-oncogene expression. Cancer Res. 62, 4169–4175.
PicoPure RNA Isolation Kit User Guide Version B, Arcturus Engineering, Mountain View, CA.
RQ1 RNase-Free DNase Technical Bulletin TB518, Promega Corporation.
Reverse Transcription System Technical Bulletin TB099, Promega Corporation.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Manning, C.B., Mossman, B.T., Taatjes, D.J. (2006). Analysis of Asbestos-Induced Gene Expression Changes in Bronchiolar Epithelial Cells Using Laser Capture Microdissection and Quantitative Reverse Transcriptase-Polymerase Chain Reaction. In: Taatjes, D.J., Mossman, B.T. (eds) Cell Imaging Techniques. Methods in Molecular Biology™, vol 319. Humana Press. https://doi.org/10.1007/978-1-59259-993-6_11
Download citation
DOI: https://doi.org/10.1007/978-1-59259-993-6_11
Publisher Name: Humana Press
Print ISBN: 978-1-58829-157-8
Online ISBN: 978-1-59259-993-6
eBook Packages: Springer Protocols