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Catalog
6. HEME I: Introduction to Technology Ondemand
C06 HEME I Intro to Technology ADubuc Notes
C06 HEME I Intro to Technology ADubuc Notes
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The document discusses the introduction to FISH (Fluorescence In Situ Hybridization) technology. FISH is a technique used to evaluate cytogenetic aberrations at a level of resolution below GTG banding. It allows for the rapid analysis of copy number and structural variations, and it eliminates the need for cell culturing. <br /><br />The document outlines the procedural steps involved in FISH, including fixation of cells, permeabilization, denaturation, hybridization, post-hybrid washing, and analysis. It also discusses the advantages of FISH-based studies, such as the ability to evaluate cytogenetic aberrations below the resolution of GTG banding and the rapid analysis of copy number and structural variations.<br /><br />The document explains different types of FISH probes, including metaphase FISH, which requires actively dividing cells, and interphase FISH, which can be used for numerical analysis but cannot reliably map signals to chromosomes. The types of probes discussed include enumeration probes, break apart probes (normal and abnormal), and dual-color, dual-fusion probes.<br /><br />Various considerations and questions related to FISH probes are discussed, such as the number of nuclei required for a reportable finding, the differentiation between gain and amplification, and the construction and overlap of probes with genes of interest.<br /><br />The document also highlights considerations during the launch and training of FISH testing in the laboratory, such as evaluating gap observed in break apart probes, ensuring concordance of FISH readers, and conducting equivalency tests between old and new lots.<br /><br />Finally, the document concludes by mentioning the benefits and limitations of FISH technology, including the ability to rapidly detect both numerical and structural aberrations, but with the limitation of only assessing the region targeted by the probes and the potential unavailability of commercial probes, which may require custom design. FISH can be performed using both FFPE and fresh material.
Keywords
FISH
Fluorescence In Situ Hybridization
cytogenetic aberrations
copy number variations
structural variations
cell culturing
metaphase FISH
interphase FISH
break apart probes
FFPE material
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