Improved technique for analysis of formalin-fixed, paraffin-embedded tumors by fluorescence in situ hybridization
| Autor: | Anne Kallioniemi, Jorma Isola, E. Hyytinen, Olli-P. Kallioniemi, Tapio Visakorpi |
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| Rok vydání: | 1994 |
| Předmět: |
Glycerol
Hot Temperature Tissue Fixation Centromere Biophysics Aneuploidy Breast Neoplasms In situ hybridization Biology Pathology and Forensic Medicine Endocrinology Formaldehyde Neoplasms medicine Chromosomes Human Humans Interphase In Situ Hybridization Fluorescence Repetitive Sequences Nucleic Acid Retrospective Studies Paraffin Embedding medicine.diagnostic_test Hybridization probe DNA–DNA hybridization Gene Amplification Chromosome DNA Neoplasm Oncogenes Cell Biology Hematology medicine.disease Proteinase K Molecular biology Chromatin Microscopy Fluorescence biology.protein DNA Probes Fluorescence in situ hybridization |
| Zdroj: | Cytometry. 16:93-99 |
| ISSN: | 1097-0320 0196-4763 |
| DOI: | 10.1002/cyto.990160202 |
| Popis: | Fluorescence in situ hybridization (FISH) and specific DNA probes for peri-centromeric repeat regions and unique sequence loci have made it possible to study chromosomal aberrations from interphase tumor nuclei. Large-scale retrospective studies on the prognostic value of interphase cytogenetics would become feasible if these techniques were readily applicable to nuclei from archival formalin-fixed tumor tissues. We describe here an improved technique for interphase FISH analysis of tumors that have been extensively fixed in formalin. The protocol aims at improving probe penetration and hybridization efficiency by inducing chromatin decondensation and swelling of the nuclei with a heat treatment in a 90 degrees C glycerol solution prior to hybridization. Using this cell pretreatment, FISH results on the detection of chromosome copy number aberrations and amplification of the c-erbB-2 oncogene from formalin-fixed, paraffin-embedded tissues were highly concordant with those from fresh tissues. In contrast to previously described methods, separate adjustments of denaturation or proteinase K digestion are not required for each sample. This method facilitates retrospective analyses of large series of tumors and is also useful for applying FISH to routine diagnostic purposes using formalin-fixed material. |
| Databáze: | OpenAIRE |
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