Method for detecting chromosomal abnormalities
A technology for chromosome and marker detection, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc., to achieve the effect of reducing sample volume and high efficiency
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Embodiment 1
[0223] Example 1: Fivefold FISH probes "SPEC ERBB2, EGFR, FGFR1, MET&SOX2FiveCheck using ZytoVision Inc. TM NG-FISH probes" FISH analysis for detection of multiple number aberrations in different cell types
[0224] FISH was performed on sections of formalin-fixed paraffin-embedded (FFPE) lung and breast tumor preparations with a thickness of 3 to 5 μm in the absence and with previously diagnosed ERBB2 gene amplification, which were applied onto a coated glass mount and bake overnight at 58°C.
[0225] To remove paraffin, the preparation was first heated on a hot plate at 70° C. for 10 minutes and then incubated in 100% xylene twice for 10 minutes at room temperature (RT). Subsequently, the preparation was rehydrated by a series of descending ethanol (5 min once, RT, in 96%, 96%, 90%, 70% denatured ethanol) and incubated in ultrapure water (twice, each two minutes, RT). To permeabilize the cells, they were subsequently heat pretreated at 98 °C for 15 min in the heat pretrea...
Embodiment 2
[0231] Example 2: FISH Analysis of Translocation of ALK and ROS1 Regions in Different Cell Types Using Quadruple FISH Probe "Zytolight SPEC ALK&ROS1 Break Apart Single-Mix NG-FISH Probe" from ZytoVision Inc.
[0232] Cells of the formalin-fixed paraffin-embedded (FFPE) Hela cell line with a thickness of 3 to 5 μm ( CCL-2 TM ), HCC78 (provided by Prof. Hildeus from Gothic) and H3122 (provided by Prof. Hildeus from Gothic) for FISH,
[0233] It was applied to a coated glass carrier and baked overnight at 58°C
[0234] To remove paraffin, the preparation was first heated on a hot plate at 70° C. for 10 minutes and then incubated in 100% xylene twice for 10 minutes at room temperature (RT). Subsequently, the modified preparations were rehydrated by descending ethanol series (5 min once, RT, in 96%, 96%, 90%, 70% denatured ethanol) and incubated in ultrapure water (twice, each two minutes, RT). To permeabilize the cells, they were subsequently heat pretreated at 98 °C for 15 m...
Embodiment 3
[0242] Example 3: FISH analysis to detect translocation of the ROS1 region in 6q22 using quadruple FISH probe "Zytolight SPECALK & ROS1 Break Apart Single-Mix NG-FISH probe" from ZytoVision, Inc.
[0243]FISH analysis of translocations in the ROS1 region at 6q22 was detected using the quadruple FISH probe "Zytolight SPEC ALK & ROS1 Break Apart SingleMix NG-FISH probe" from ZytoVision, Inc. The probe is a mixture based on four site-specific hybridization probes consisting of a green-labeled polynucleotide (absorbing at 503nm and emitting at 528nm) targeting the ALK breakpoint region in 2p23 Sequences nearby, and in 6q22 for sequences located near the ROS1 breakpoint region, orange labeled polynucleotides (absorbing at 547 nm and emitting at 572 nm), which in 2p23 for sequences located distal to the ALK breakpoint region , and consisted of blue-labeled polynucleotides (absorbing at 426 nm and emitting at 480 nm) in 6q22 for sequences located distal to the ROS1 breakpoint region,...
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