Abstract
Determining how a molecular targeting agent affects signaling pathways is challenging. To facilitate this exploration, we use a combination of medium-throughput methodologies such as tissue microarrays (TMA) and digital image analysis for quantification of immunohistochemistry (IHC). Because TMAs enable the simultaneous analysis of up to 1000 tissues, this tool can be used to comprehensively assess changes in signal transduction pathways over time. TMAs can be digitized and analyzed using the Aperio ScanScope imaging system and Aperio ImageScope software to deliver semiquantitative data. This chapter describes the methodologies for tissue collection and for the construction, sectioning, staining, and digital analysis of TMAs.
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References
Kumar B, De Silva M, Venter DJ et al (2004) Tissue microarrays: a practical guide. Pathology 36(4):295–300
Watanabe A, Cornelison R, Hostetter G (2005) Tissue microarrays: applications in genomic research. Expert Rev Mol Diagn 5(2):171–181
Kononen J, Bubendorf L, Kallioniemi A et al (1998) Tissue microarrays for high-throughput molecular profiling of tumor specimens. Nat Med 4(7):844–847
Kallioniemi OP, Wagner U, Kononen J et al (2001) Sauter G: tissue microarray technology for high-throughput molecular profiling of cancer. Hum Mol Genet 10(7):657–662
Alvarenga AW, Coutinho-Camillo CM, Rodrigues BR et al (2013) A comparison between manual and automated evaluations of tissue microarray patterns of protein expression. J Histochem Cytochem 61(4):272–282
Kalra J, Dragowska WH, Bally MB (2015) Using pharmacokinetic profiles and digital quantification of stained tissue microarrays as a medium-throughput, quantitative method for measuring the kinetics of early signaling changes following integrin-linked kinase inhibition in an in vivo model of cancer. J Histochem Cytochem 63(9):691–709
Lavorato-Rocha AM, Anjos LG, Cunha IW et al (2015) Immunohistochemical assessment of PTEN in vulvar cancer: best practices for tissue staining, evaluation, and clinical association. Methods 77–78:20–24
Rexhepaj E, Jirstrom K, O'Connor DP et al (2010) Validation of cytoplasmic-to-nuclear ratio of survivin as an indicator of improved prognosis in breast cancer. BMC Cancer 10:639
Kokkat TJ, Patel MS, McGarvey D et al (2013) Archived formalin-fixed paraffin-embedded (FFPE) blocks: a valuable underexploited resource for extraction of DNA, RNA, and protein. Biopreserv Biobank 11(2):101–106
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Kalra, J., Baker, J. (2017). Using Digital Quantification of Stained Tissue Microarrays as a Medium-Throughput, Quantitative Method for Measuring the Kinetics of Signal Transduction. In: Kalyuzhny, A. (eds) Signal Transduction Immunohistochemistry. Methods in Molecular Biology, vol 1554. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6759-9_5
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DOI: https://doi.org/10.1007/978-1-4939-6759-9_5
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6757-5
Online ISBN: 978-1-4939-6759-9
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