title: Fluorescent In Situ Hybridization in Suspension by Imaging Flow Cytometry.
abstract: The emergence of imaging flow cytometry (IFC) has brought novel applications exploiting its advantages over conventional flow cytometry and microscopy. One of the new applications is fluorescence in situ hybridization in suspension (FISH-IS). Conventional FISH is a slide-based approach in which the spotlike imagery resulting from hybridization with fluorescently tagged probes is evaluated by fluorescence microscopy. The FISH-IS approach evaluated by IFC enables the evaluation of tens to hundreds of thousands of cells in suspension and the analysis can be automated and standardized diminishing operator bias from the analysis. The high cell number throughput of FISH-IS improves the detection of rare events compared to conventional FISH. The applicability of FISH-IS is currently limited to detection of abnormal quantitative differences of hybridization targets such as occur in numerical chromosome abnormalities, deletions and amplifications.Here, we describe a protocol for FISH-IS using chromosome enumeration probes as an example.
authors.affiliation: Department of Flow and Image Cytometry, Roswell Park Cancer Institute, Buffalo, NY, 14263, USA. Orla.Maguire@RoswellPark.org.
authors.forename: Orla
authors.initials: O
authors.lastname: Maguire
authors.affiliation: Department of Flow and Image Cytometry, Roswell Park Cancer Institute, Buffalo, NY, 14263, USA.
authors.forename: Paul K
authors.initials: PK
authors.lastname: Wallace
authors.affiliation: Department of Flow and Image Cytometry, Roswell Park Cancer Institute, Buffalo, NY, 14263, USA.
authors.forename: Hans
authors.initials: H
authors.lastname: Minderman
mesh_terms: D002869:Chromosome Aberrations; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007963:Leukocytes, Mononuclear
keywords: Acute myeloid leukemia; Cytogenetic abnormalities; FISH in suspension; ImageStream