Abstract
The technique of nucleic acid in situ hybridization is an effective method for identifying the existence and abundance of nucleic acids in tissue sections or cytological preparations. Such a method has the advantage of keeping morphological relationships intact while identifying changes at the molecular level. As a noncoding regulatory RNA, microRNA has been found to intricately control many physiological and pathological conditions. We provide here a representative fluorescence in situ hybridization protocol for microRNA detection, and note commonly used alternatives, and some troubleshooting points. The method described is based on formalin-fixed paraffin-embedded oral cancer tissues but should be broadly applicable to similarly processed tissues of other types of cancer.
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References
Lee RC, Feinbaum RL, Ambros V (1993) The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 75:843–854
Andorfer CA, Necela BM, Thompson EA, Perez EA (2011) MicroRNA signatures: clinical biomarkers for the diagnosis and treatment of breast cancer. Trends Mol Med 17:313–319
Qavi AJ, Kindt JT, Bailey RC (2010) Sizing up the future of microRNA analysis. Anal Bioanal Chem 398:2535–2549
Nielsen BS (2012) MicroRNA in situ hybridization. Methods Mol Biol 822:67–84
Shi Z, Johnson JJ, Stack MS (2012) Fluorescence in situ hybridization for microRNA detection in archived oral cancer tissues. J Oncol 2012:903581
McKay JA, Murray GI, Keith WN, McLeod HL (1997) Amplification of fluorescent in situ hybridisation signals in formalin fixed paraffin wax embedded sections of colon tumour using biotinylated tyramide. Mol Pathol 50:322–325
Bobrow MN, Moen PT Jr (2001) Tyramide signal amplification (TSA) systems for the enhancement of ISH signals in cytogenetics. Curr Protoc Cytom Chapter 8, Unit 8 9
Helander KG (1994) Kinetic studies of formaldehyde binding in tissue. Biotech Histochem 69:177–179
Fox CH, Johnson FB, Whiting J, Roller PP (1985) Formaldehyde fixation. J Histochem Cytochem 33:845–853
Glud M, Klausen M, Gniadecki R, Rossing M, Hastrup N, Nielsen FC et al (2009) MicroRNA expression in melanocytic nevi: the usefulness of formalin-fixed, paraffin-embedded material for miRNA microarray profiling. J Invest Dermatol 129:1219–1224
Liu A, Tetzlaff MT, Vanbelle P, Elder D, Feldman M, Tobias JW et al (2009) MicroRNA expression profiling outperforms mRNA expression profiling in formalin-fixed paraffin-embedded tissues. Int J Clin Exp Pathol 2:519–527
Nuovo GJ (2010) In situ detection of microRNAs in paraffin embedded, formalin fixed tissues and the co-localization of their putative targets. Methods 52:307–315
Kloosterman WP, Wienholds E, de Bruijn E, Kauppinen S, Plasterk RH (2006) In situ detection of miRNAs in animal embryos using LNA-modified oligonucleotide probes. Nat Methods 3:27–29
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Shi, Z., Johnson, J.J., Stack, M.S. (2013). Detecting MicroRNA in Human Cancer Tissues with Fluorescence In Situ Hybridization. In: Kolpashchikov, D., Gerasimova, Y. (eds) Nucleic Acid Detection. Methods in Molecular Biology, vol 1039. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-535-4_2
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DOI: https://doi.org/10.1007/978-1-62703-535-4_2
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-534-7
Online ISBN: 978-1-62703-535-4
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