Abstract
Circulating tumor cells (CTCs) may represent an easily accessible source of tumor material to assess genetic aberrations such as gene-rearrangements or gene-amplifications and screen cancer patients eligible for targeted therapies. As the number of CTCs is a critical parameter to identify such biomarkers, we developed fluorescent in situ hybridization (FISH) for CTCs enriched on filters (filter-adapted-FISH, FA-FISH). Here, we describe the FA-FISH protocol, the combination of immunofluorescent staining (DAPI/CD45) and FA-FISH techniques, as well as the semi-automated microscopy method that we developed to improve the feasibility and reliability of FISH analyses in filtration-enriched CTC.
Keywords:
Circulating tumor cells; FA-FISH; FISH; Filtration-enrichment; ISET.
MeSH terms
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Antibodies, Monoclonal / chemistry
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Antibodies, Monoclonal / metabolism
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Biomarkers, Tumor / genetics
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Biomarkers, Tumor / immunology
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Biomarkers, Tumor / metabolism
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Cell Count
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Cell Separation / instrumentation
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Cell Separation / methods*
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Cell Size
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Equipment Design
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Filtration / instrumentation
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Filtration / methods*
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Fluorescent Antibody Technique / methods
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Fluorescent Dyes / chemistry
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Humans
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Immunoconjugates / chemistry
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In Situ Hybridization, Fluorescence / methods*
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Indoles / chemistry
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Leukocyte Common Antigens / genetics
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Leukocyte Common Antigens / immunology
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Leukocyte Common Antigens / metabolism
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Neoplasms / blood
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Neoplasms / diagnosis*
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Neoplasms / immunology
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Neoplasms / pathology
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Neoplastic Cells, Circulating / immunology
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Neoplastic Cells, Circulating / metabolism
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Neoplastic Cells, Circulating / pathology*
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Protein Binding
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Rheology
Substances
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Antibodies, Monoclonal
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Biomarkers, Tumor
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Fluorescent Dyes
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Immunoconjugates
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Indoles
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DAPI
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Leukocyte Common Antigens
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PTPRC protein, human