title: Simultaneous imaging of membrane antigen and the corresponding chromosomal locus in pathology archives.
abstract: A new procedure for the simultaneous staining of membranous antigens, such as tyrosine kinase-type cell surface receptor HER2 (c-erbB2), and the corresponding chromosome (chromosome 17 for c-erbB2) in the same cell for use in examining pathology archives is presented. A multistep procedure involving microwave-assisted fluorescence in situ hybridization and immunofluorescence yielded cell images having c-erbB2 on the membrane and genomic signals from the chromosome 17 centromere and the c-erbB2 locus. Furthermore, a combination of microwave-assisted chromogenic in situ hybridization and immunohistochemistry found colorized signals from both chromosome 17 centromere in the nuclei and c-erbB2 on the membranes of individual cells. Quantitative image analysis further confirmed the presence of a significantly stronger c-erbB2 immunoreactivity on cells containing three or more signals from chromosome 17 than from those with less than three signals. It was possible to extend the constellation of cell surface markers and corresponding chromosomes or locus-specific makers to several other genes including CDH1. In this case, the disappearances of CDH1 expression, a CDH1 locus signal, and a centromere enumeration probe (CEP) 16 signal were simultaneously demonstrated in the less-adhesive tumor cells. Thus, it is believed that this procedure might pave the way for exploiting pathology archives for the genotype-phenotype analysis of individual cells.
authors.affiliation: Department of Pathology, Hamamatsu University of Medicine, Hamamatsu, Japan.
authors.forename: Hisaki
authors.initials: H
authors.lastname: Igarashi
authors.forename: Kimihiro
authors.initials: K
authors.lastname: Yamashita
authors.forename: Masaya
authors.initials: M
authors.lastname: Suzuki
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Kitayama
authors.forename: Jun
authors.initials: J
authors.lastname: Isogaki
authors.forename: Keiji
authors.initials: K
authors.lastname: Maruyama
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Sunayama
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Tsuda
authors.forename: Takachika
authors.initials: T
authors.lastname: Ozawa
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Kiyose
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Sugimura
mesh_terms: D000954:Antigens, Surface / Q000235:genetics* / Q000378:metabolism; D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002503:Centromere; D002885:Chromosomes, Human, Pair 16; D002886:Chromosomes, Human, Pair 17; D015342:DNA Probes; D005260:Female; D005838:Genotype; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008872:Microwaves; D010641:Phenotype; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism
chemical_list: D000954:Antigens, Surface; D015820:Cadherins; D015342:DNA Probes; D018719:Erb-b2 Receptor Tyrosine Kinases