Abstract
RNA in situ hybridization (ISH) complements immunohistochemistry (IHC) by confirming localized gene expression in a tissue. Effective use of ISH requires careful communication and coordination between molecular biologists, histologists, and pathologists. This chapter reviews the literature and experience of the authors in the use of formalin-fixed paraffin-embedded tissues for RNA histology techniques.
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References
Eick D et al (1985) Aberrant c-myc RNAs of Burkitt’s lymphoma cells have longer half-lives. EMBO J 4(13B):3717
Gall JG, Pardue ML (1969) Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc Natl Acad Sci 63(2):378–383
John H, Birnstiel M, Jones K (1969) RNA-DNA hybrids at the cytological level. Nature 223(5206):582–587
Femino AM et al (1998) Visualization of single RNA transcripts in situ. Science 280(5363):585–590
Raj A et al (2008) Imaging individual mRNA molecules using multiple singly labeled probes. Nat Methods 5(10):877–879
Kitaoka M et al (2012) Transglutaminase-mediated in situ hybridization (TransISH) system: a new methodology for simplified mRNA detection. Anal Chem 84(14):5885–5891
Adams J (1992) Biotin amplification of biotin and horseradish peroxidase signals in histochemical stains. J Histochem Cytochem 40(10):1457–1463
Chen CH et al (2011) Specific sorting of single bacterial cells with microfabricated fluorescence-activated cell sorting and tyramide signal amplification fluorescence in situ hybridization. Anal Chem 83(19):7269–7275
Larsson C et al (2010) In situ detection and genotyping of individual mRNA molecules. Nat Methods 7(5):395–397
Nilsson M (2006) Lock and roll: single-molecule genotyping in situ using padlock probes and rolling-circle amplification. Histochem Cell Biol 126(2):159–164
Player AN et al (2001) Single-copy gene detection using branched DNA (bDNA) in situ hybridization. J Histochem Cytochem 49(5):603–611
Bagasra O (2007) Protocols for the in situ PCR-amplification and detection of mRNA and DNA sequences. Nat Protoc 2(11):2782–2795
Komminoth P (1992) Digoxigenin as an alternative probe labeling for in situ hybridization. Diagn Mol Pathol 1(2):142–150
Song R, Ro S, Yan W (2010) In situ hybridization detection of microRNAs. In: RNA therapeutics. Springer, pp 285–292
Jørgensen S et al (2010) Robust one-day in situ hybridization protocol for detection of microRNAs in paraffin samples using LNA probes. Methods 52(4):375–381
Evans MF, Aliesky HA, Cooper K (2003) Optimization of biotinyl-tyramide-based in situ hybridization for sensitive background-free applications on formalin-fixed, paraffin-embedded tissue specimens. BMC Clin Pathol 3(1):2
Lambros MB et al (2006) Unlocking pathology archives for molecular genetic studies: a reliable method to generate probes for chromogenic and fluorescent in situ hybridization. Lab Invest 86(4):398–408
Wilcox JN (1993) Fundamental principles of in situ hybridization. J Histochem Cytochem 41(12):1725–1733
Zirbel R et al (1993) Evidence for a nuclear compartment of transcription and splicing located at chromosome domain boundaries. Chromosome Res 1(2):93–106
Niedobitek G et al (1989) Detection of viral genomes in the liver by in situ hybridization using 35S-, bromodeoxyuridine-, and biotin-labeled probes. Am J Pathol 134(3):633
Garrett KL, Grounds MD, Beilharz MW (1992) Nonspecific binding of nucleic acid probes to Paneth cells in the gastrointestinal tract with in situ hybridization. J Histochem Cytochem 40(10):1613–1618
Steiner I et al (1989) Effects of lipofuscin on in situ hybridization in human neuronal tissue. J Virol Methods 24(1):1–9
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Potts, S.J., Peljto, M., Suraneni, M., Krueger, J.S. (2014). Measuring the Messenger: RNA Histology in Formalin-Fixed Tissues. In: Potts, S., Eberhard, D., Wharton, Jr., K. (eds) Molecular Histopathology and Tissue Biomarkers in Drug and Diagnostic Development. Methods in Pharmacology and Toxicology. Humana Press, New York, NY. https://doi.org/10.1007/7653_2014_19
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DOI: https://doi.org/10.1007/7653_2014_19
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