Discrimination of distinct subpopulations within a tumor with combined double immunophenotyping and interphase cytogenetics.

Weber‐Matthiesen, Klaus; Müller-Hermelink, A; Deerberg, J.; Scherthan, Harry; Schlegelberger, Brigitte; Grote, W.

doi:10.1177/41.11.7691932

Cited by 19 publications

(6 citation statements)

References 7 publications

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“…The FICTION technique was developed to assign tumor cells to a cytogenetically defined clone and, at the same time, to determine their specific cell lineage (Weber-Matthiesen et al1992,1993. FICTION has been proven to be clinically meaningful in hematological neoplasms (Nylund et al 1993;Haferlach et al 1997;Temple et al 2004;Young et al 2006).…”

Section: Discussionmentioning

confidence: 99%

Phenotypic and Genetic Characterization of Circulating Tumor Cells by Combining Immunomagnetic Selection and FICTION Techniques

Campos

Prior²,

Warleta

et al. 2008

J Histochem Cytochem.

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S U M M A R Y The presence of circulating tumor cells (CTCs) in breast cancer patients has been proven to have clinical relevance. Cytogenetic characterization of these cells could have crucial relevance for targeted cancer therapies. We developed a method that combines an immunomagnetic selection of CTCs from peripheral blood with the fluorescence immunophenotyping and interphase cytogenetics as a tool for investigation of neoplasm (FICTION) technique. Briefly, peripheral blood (10 ml) from healthy donors was spiked with a predetermined number of human breast cancer cells. Nucleated cells were separated by double density gradient centrifugation of blood samples. Tumor cells (TCs) were immunomagnetically isolated with an anti-cytokeratin antibody and placed onto slides for FICTION analysis. For immunophenotyping and genetic characterization of TCs, a mixture of primary monoclonal anti-pancytokeratin antibodies was used, followed by fluorescent secondary antibodies, and finally hybridized with a TOP2A/HER-2/CEP17 multicolor probe. Our results show that TCs can be efficiently isolated from peripheral blood and characterized by FICTION. Because genetic amplification of TOP2A and ErbB2 (HER-2) in breast cancer correlates with response to anthracyclines and herceptin therapies, respectively, this novel methodology could be useful for a better classification of patients according to the genetic alterations of CTCs and for the application of targeted therapies. (J Histochem Cytochem 56:667-675, 2008)

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Section: Discussionmentioning

confidence: 99%

Phenotypic and Genetic Characterization of Circulating Tumor Cells by Combining Immunomagnetic Selection and FICTION Techniques

Campos

Prior²,

Warleta

et al. 2008

J Histochem Cytochem.

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“…There are now definite clinical indications for the application of molecular cytogenetic techniques in the diagnosis of several types of non-CNS cancer, for example, there is a correlation between numerical abnormalities of chromosomes 11 and 17 and lymph node metastasis of primary breast cancer [27] and collaboration between scientists and clinicians will rapidly define the areas in which these techniques will be of clinical utility in diagnostic neurooncology. Similarly, hybrid techniques which allow for simultaneous immunophenotyping and interphase cytogenetic analysis (for example, the FICTION method [82,83]), which have been applied to lymphomas are likely to be applied to cells from solid tumours and which will provide a further spur to the acceptance of these types of technique to routine neuropathology.…”

Section: Discussionmentioning

confidence: 99%

Non‐isotopic molecular cytogenetics in neuro‐oncology

Darling

Warr

Ashmore

et al. 1997

Neuropathology Appl Neurobio

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The molecular genetic analysis of brain tumours has been the focus of considerable interest for a number of years. However, these studies have been largely directed towards understanding the fundamental biological processes involved in tumorigenesis and the techniques which have been used require considerable molecular biological skills. Unfortunately, there has not been the impetus to correlate basic biological studies with clinical or neuropathological features. The development of non-isotopic molecular cytogenetic in situ hybridization (ISH) techniques which can be applied to archival tumour material provides an opportunity to address a wide range of neuropathological questions at a genetic level. Identification of specific chromosomes has been made possible by the isolation of probes which recognize the highly repeated sequences present in the centromeric regions of individual chromosomes. Libraries of human chromosome-specific painting probes are also available. A range of probes which bind to the whole or part of specific single copy genes are becoming available. These can be detected with either fluorochromes with different emission colours or with enzymatic detection systems in either interphase nuclei derived from fresh, fixed and embedded tumour samples, touch preparations or smears (so-called 'interphase cytogenetics') as well as conventional metaphase spreads. Comparative genomic hybridization can be used to scan the entire genome for deletions or amplifications without any pre-existing information about the likely locations of these abnormalities or the availability of any specific DNA probes. These techniques can be used to identify aneuploidy or structural alterations in individual chromosomes and are likely to yield important information about the location of genes important in the pathogenesis of brain tumours and may also provide the basis for the refinement of diagnostic or prognostic criteria of these neoplasms.

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“…integrated immunofluorescent labeling of cellular antigens and FISH, or “immunoFISH.” “Fluorescence immunophenotype and interphase cytogenetics as a tool for investigation of neoplasms” or “FICTION” is a powerful tool for the identification of genetic abnormalities in cells based on phenotype which can be performed on cytocentrifuge preparations or tissue sections to detect specific genetic abnormalities in cells identified by their phenotype (e.g. CD138‐positive plasma cells) . This also enables primary genetic abnormalities present in all neoplastic cells (e.g.…”

Section: Fish In Suspensionmentioning

confidence: 99%

Development of a robust immuno‐S‐FISH protocol using imaging flow cytometry

et al. 2016

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Fluorescence in situ hybridization (FISH) is a microscopy technique which uses a fluorescent probe to detect DNA sequences and is generally performed on metaphase spreads or interphase nuclei of intact cells on a slide. In a diagnostic laboratory, cells are hybridized with fluorescent probes and up to 200 cells counted for the number of cells with probe "spots." Recent modifications to standard FISH include immuno-FISH, where chromosomal abnormalities are detected only in cells by their phenotype, and S-FISH where probe hybridization is performed on whole cells in suspension. Here we describe the development of an immuno-S-FISH method that combines immunophenotyping and FISH analysis of cells in suspension followed by analysis on an imaging flow cytometer. This single platform technique couples microscopy with flow cytometry and "spot" detection of bound FISH probe. Automated immuno-S-FISH enables large numbers of analyzed cells to be identified by phenotype and assessed for specific chromosomal determinants by FISH. This novel robust method enables quantitative cell population analysis and "spot" counting for large numbers of cells. We report method optimization of this imaging immuno-S-FISH flow cytometry protocol which has capability for many clinical applications. V C 2016 International Society for Advancement of Cytometry Key terms imaging cytometry; S-FISH; immunophenotyping; hematological malignancy CYTOGENETIC analysis is an integral component in the assessment of many constitutional and neoplastic disorders, particularly those of hemopoietic origin. This has traditionally been performed by full chromosomal analysis (karyotyping) which provides a global analysis of the entire genome; however, this has limitations due to low resolution and the need to have cells in metaphase of the cell cycle. Fluorescence in situ hybridization (FISH) is more sensitive than karyotyping in identifying specific genomic defects and can be performed in nondividing cells (1,2). FISH is based on fluorescently-labeled single-stranded DNA probe annealing to its complementary sequence in a target genome to detect DNA sequences and submicroscopic genetic changes (3). The probes are designed to target genes of interest to identify rearrangements, deletions, and gains in both whole cells in interphase and metaphase. The probes can be whole chromosome paints or locus-specific and these bind to the parts of the chromosome with which they show a high degree of sequence homology. Locus-specific probes are most frequently used and are of three main types:1. Copy number probes: centromeric probes are used to detect loss or gain of whole chromosomes and ploidy changes. These are used to enumerate specific chromosomes in interphase cells, providing an accurate method to ascertain chromosome copy number (4). 2. Breakapart probes (e.g. MLL locus located to chromosome band 11q23) (5) 3. Dual-fusion probes (e.g. BCR-ABL1 fusion gene resulting from t(9;22)(q34;q11)) (6,7)

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Discrimination of distinct subpopulations within a tumor with combined double immunophenotyping and interphase cytogenetics.

Cited by 19 publications

References 7 publications

Phenotypic and Genetic Characterization of Circulating Tumor Cells by Combining Immunomagnetic Selection and FICTION Techniques

Phenotypic and Genetic Characterization of Circulating Tumor Cells by Combining Immunomagnetic Selection and FICTION Techniques

Non‐isotopic molecular cytogenetics in neuro‐oncology

Development of a robust immuno‐S‐FISH protocol using imaging flow cytometry

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