title: Analysis of receptor tyrosine kinase gene amplification on the example of FGFR1.
abstract: FISH (fluorescent in situ hybridization) is a molecular cytogenetic method to detect large-scale genetic alterations in tissue and/or cells. Numerical aberrations (deletions and amplifications) and structural aberrations (translocations and fusions) are detectable. Probes bind complementary to the DNA strand of the region of interest. Subsequently, the probes are detected via fluorochromes and appear as colored dots that can be assessed under the fluorescence microscope.In situ hybridization is divided into three steps: pretreatment, hybridization, and posthybridization. Pretreatment opens up the cell membranes for hybridization, so that the probe can bind to the complementary DNA target. Posthybridization includes washing steps to remove excessive probes and detection of probes via secondary marked fluorochromes. DAPI stains nuclei and serves as mounting media.
authors.affiliation: Department of Prostate Cancer Research, Institute of Pathology, University Hospital of Bonn, Sigmund-Freud-Straße 25, 53127, Bonn, Germany.
authors.forename: Diana
authors.initials: D
authors.lastname: Boehm
authors.forename: Anne
authors.initials: A
authors.lastname: von Mässenhausen
authors.forename: Sven
authors.initials: S
authors.lastname: Perner
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D022202:Chromosomes, Artificial, Bacterial / Q000737:chemistry / Q000378:metabolism; D015342:DNA Probes / Q000138:chemical synthesis / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005784:Gene Amplification; D015870:Gene Expression; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008214:Lymphocytes / Q000378:metabolism* / Q000648:ultrastructure; D008677:Metaphase; D009693:Nucleic Acid Hybridization; D010539:Permeability; D051496:Receptor, Fibroblast Growth Factor, Type 1 / Q000235:genetics* / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D015342:DNA Probes; C496348:FGFR1 protein, human; D051496:Receptor, Fibroblast Growth Factor, Type 1