Improved technique for analysis of formalin-fixed, paraffin-embedded tumors by fluorescence in situ hybridization

Journal Article · Wed Jun 01 04:00:00 EDT 1994 · Cytometry

DOI:https://doi.org/10.1002/cyto.990160202· OSTI ID:114858

Hyytinen, Eija; Visakorpi, T; Kallioniemi, O P ^[1]

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1. Tampere Univ. Hospital (Finland); and others

Fluorescence in situ hybridization (FISH) and specific DNA probes for pericentromeric 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. The authors 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 of proteinase K digestion are not required for each sample. This method facilitates retrospective analyses of large species of tumors and is also useful for applying FISH to routine diagnostic purposes using formalin-fixed material. 32 refs., 1 fig., 2 tabs.

• Sponsoring Organization: USDOE
• OSTI ID: 114858
• Journal Information: Cytometry, Journal Name: Cytometry Journal Issue: 1-4 Vol. 16; ISSN CYTODQ; ISSN 0196-4763
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
CHROMOSOMAL ABERRATIONS
DETECTION
DNA HYBRIDIZATION
NEOPLASMS