c-MYC and centromere 8 copy number status of MCF-10A cell populations was determined by dual hybridisation with a Vysis LSI SpectrumOrange c-MYC Probe (Abbott Molecular, Maidenhead, UK; cat. no.: 05J545-011) and a CEP8 SpectrumGreen Probe (Abbott Molecular; cat. no.: 06J37-018) as recommended by the suppliers. c-MYC (orange), centromere 8 (aqua) and IGH (green) copy number status of HuMECs was determined using the Abbott IGH/MYC/CEP8 Tri-colour Dual Fusion Translocation Probe (Abbott Molecular). Slides were heated to 72 °C for 5 min and then incubated for 24 h at 37 °C in a humidified hybridisation chamber (HYBrite; Abbott Molecular). After hybridisation, slides were counterstained with 4′,6-diamidino-2-phenylindole (Vector Laboratories, Peterborough, UK). FISH was scored with an Olympus BX-61 fluorescence microscope (Olympus, Southend-on-Sea, UK) with a ×100 oil objective. Images were analysed using the CytoVision 7.2 SPOT counting system (Leica Microsystems, Gateshead, UK). A minimum of 100 (for MCF-10A) or 70 (for HuMECs) nuclei were scored per test by two independent analysts.
Cytovision 7.2 spot counting system
Manufactured by Leica
Sourced in United Kingdom
The CytoVision 7.2 SPOT counting system is a laboratory equipment product designed for cell counting and analysis. It provides automated detection and enumeration of cells or particles in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Hybrite, by Abbott (3 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (3 mentions)
Bx61 fluorescence microscope, by Olympus (3 mentions)
Vysis lsi spectrumorange c myc probe, by Abbott (2 mentions)
Cep8 spectrumgreen probe, by Abbott (2 mentions)
Igh myc cep8 tri colour dual fusion translocation probe, by Abbott (2 mentions)
3 protocols using cytovision 7.2 spot counting system
1
Fluorescent In Situ Hybridization Assay for c-MYC and Centromere 8 in MCF-10A and HuMECs
2
MYC Breakapart FISH Analysis
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FISH analysis was performed by the Cancer Cytogenetics department at Newcastle Hospitals NHS Foundation Trust, according to established protocols. The Cytocell MYC ‘breakapart’ Probe set was hybridised to nuclei as recommended by the suppliers. Slides were heated to 72°C for 5 min and then incubated for 24 hr at 37°C in a humidified hybridisation chamber (HYBrite; Abbott Molecular). After hybridisation, slides were counterstained with 4′,6-diamidino-2-phenylindole (Vector Laboratories, Peterborough, UK). FISH was scored with an Olympus BX-61 fluorescence microscope (Olympus, Southend-on-Sea, UK) with a × 100 oil objective. Images were analysed using the CytoVision 7.2 SPOT counting system (Leica Microsystems, Gateshead, UK). A minimum of 100 nuclei were scored per test by two independent analysts.
3
Fluorescent in situ Hybridization Analysis of c-MYC and Centromere 8 in Breast Cell Lines
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c-MYC and centromere 8 copy number status of MCF-10A cell populations was determined by dual hybridisation with a Vysis LSI SpectrumOrange c-MYC Probe (Abbott Molecular, Maidenhead, UK; cat. no.: 05J545-011) and a CEP8 SpectrumGreen Probe (Abbott Molecular; cat. no.: 06J37-018) as recommended by the suppliers. c-MYC (orange), centromere 8 (aqua) and IGH (green) copy number status of HuMECs was determined using the Abbott IGH/MYC/CEP8 Tri-colour Dual Fusion Translocation Probe (Abbott Molecular). Slides were heated to 72 °C for 5 min and then incubated for 24 h at 37 °C in a humidified hybridisation chamber (HYBrite; Abbott Molecular). After hybridisation, slides were counterstained with 4′,6-diamidino-2-phenylindole (Vector Laboratories, Peterborough, UK). FISH was scored with an Olympus BX-61 fluorescence microscope (Olympus, Southend-on-Sea, UK) with a x100 oil objective. Images were analysed using the CytoVision 7.2 SPOT counting system (Leica Microsystems, Gateshead, UK). A minimum of 100 (for MCF-10A) or 70 (for HuMECs) nuclei were scored per test by two independent analysts.
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