ArticleMolecular pathology : MP2003
A simple and reliable pretreatment protocol facilitates fluorescent in situ hybridisation on tissue microarrays of paraffin wax embedded tumour samples.
Article in Molecular pathology : MP, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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27 citing papers in PubMed, 96 citations in OpenAlex.
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- Fast fluorescence in situ hybridisation for the enhanced detection of MET in non-small cell lung cancer.PloS one · 2019Article
- One-fits-all pretreatment protocol facilitating Fluorescence In Situ Hybridization on formalin-fixed paraffin-embedded, fresh frozen and cytological slides.Molecular cytogenetics · 2019Article
- Fluorescence In Situ Hybridization of Cells, Chromosomes, and Formalin-Fixed Paraffin-Embedded Tissues.Methods in molecular biology (Clifton, N.J.) · 2017Article
- Propagation of pure fetal and maternal mesenchymal stromal cells from terminal chorionic villi of human term placenta.Scientific reports · 2015Article
- Quantitative multigene FISH on breast carcinomas identifies der(1;16)(q10;p10) as an early event in luminal A tumors.Genes, chromosomes & cancer · 2015Article
- Prognostic impact of PHIP copy number in melanoma: linkage to ulceration.The Journal of investigative dermatology · 2014Article
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- Tandem duplication of chromosomal segments is common in ovarian and breast cancer genomes.The Journal of pathology · 2012Article
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- Pleckstrin homology domain-interacting protein (PHIP) as a marker and mediator of melanoma metastasis.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
- Validation of the histone methyltransferase EZH2 as a therapeutic target for various types of human cancer and as a prognostic marker.Cancer science · 2011Article
- Minichromosome Maintenance Protein 7 is a potential therapeutic target in human cancer and a novel prognostic marker of non-small cell lung cancer.Molecular cancer · 2011Article
- Progressive 3q amplification consistently targets SOX2 in preinvasive squamous lung cancer.American journal of respiratory and critical care medicine · 2010Article
- DNA copy number loss and allelic imbalance at 2p16 in lung cancer associated with asbestos exposure.British journal of cancer · 2009Article
- Article
Corrections and comments
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Authors and funding
10 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
aimsTo describe a robust pretreatment protocol for preparing paraffin wax embedded tissues on tissue microarrays for fluorescence in situ hybridisation (FISH). The newly developed pretreatment protocol described here was compared with the commonly used sodium thiocyanate based protocol and two different heating methods used in standard antigen unmasking protocols for immunohistochemistry (pressure cooking and microwaving in citrate acid buffer).
methodsDewaxed tissue sections were incubated in 10mM citric acid buffer at 80 degrees C for 30 minutes to two hours, followed by a short pepsin digestion (1-5 mg/ml). Pretreated tissues were co-denatured with DNA probes at 80 degrees C for 10 minutes, followed by hybridisation at 37 degrees C for 48-72 hours.
resultsThe three protocols using citrate acid buffer produced FISH signals with superior signal to noise ratios compared with sodium thiocyanate pretreatment. Most importantly, the best tissue attachment was achieved using the newly developed pretreatment protocol: on tissue microarrays less than 1% of cores were lost. To date, a total of 30 probes have been successfully hybridised on to breast tissue and multi-tissue microarrays.
conclusionThis pretreatment protocol is easy, reproducible, and facilitates FISH on tissue microarrays, with potential for widespread application in cancer research.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.