title: Image cytometry-based detection of aneuploidy by fluorescence in situ hybridization in suspension.
abstract: Cytogenetic abnormalities are important diagnostic and prognostic criteria for hematologic malignancies. Karyotyping and fluorescence in situ hybridization (FISH) are the conventional methods by which these abnormalities are detected. The sensitivity of these microscopy-based methods is limited by the abundance of the abnormal cells in the samples and therefore these analyses are commonly not applicable to minimal residual disease (MRD) stages. A flow cytometry-based imaging approach was developed to detect chromosomal abnormalities following FISH in suspension (FISH-IS), which enables the automated analysis of several log-magnitude higher number of cells compared with the microscopy-based approaches. This study demonstrates the applicability of FISH-IS for detecting numerical chromosome aberrations, establishes accuracy, and sensitivity of detection compared with conventional FISH, and feasibility to study procured clinical samples of acute myeloid leukemia (AML). Male and female healthy donor peripheral blood mononuclear cells hybridized with combinations of chromosome enumeration probes (CEP) 8, X, and Y served as models for disomy, monosomy, and trisomy. The sensitivity of detection of monosomies and trisomies amongst 20,000 analyzed cells was determined to be 1% with a high level of precision. A high correlation (R(2) = 0.99) with conventional FISH analysis was found based on the parallel analysis of diagnostic samples procured from 10 AML patients with trisomy 8 (+8). Additionally, FISH-IS analysis of samples procured at the time of clinical remission demonstrated the presence of residual +8 cells indicating that this approach may be used to detect MRD and associated chromosomal defects.
authors: Flow and Image Cytometry Laboratory, Department of Pathology and Laboratory Medicine, Roswell Park Cancer Institute, Buffalo, New York 14263, USA. hans.minderman@roswellpark.org
authors: Hans
authors: H
authors: Minderman
authors: Kristen
authors: K
authors: Humphrey
authors: Jane K
authors: JK
authors: Arcadi
authors: Andrzej
authors: A
authors: Wierzbicki
authors: Orla
authors: O
authors: Maguire
authors: Eunice S
authors: ES
authors: Wang
authors: AnneMarie W
authors: AW
authors: Block
authors: Sheila N J
authors: SN
authors: Sait
authors: Thaddeus C
authors: TC
authors: George
authors: Paul K
authors: PK
authors: Wallace
mesh_terms: D000465:Algorithms; D000782:Aneuploidy; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007091:Image Processing, Computer-Assisted; D017404:In Situ Hybridization, Fluorescence; D015470:Leukemia, Myeloid, Acute / Q000235:genetics / Q000473:pathology*; D007963:Leukocytes, Mononuclear / Q000473:pathology*; D057230:Limit of Detection; D008297:Male; D008954:Models, Biological; D015203:Reproducibility of Results; D059010:Single-Cell Analysis