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Introduction
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ZytoDot ® products are designed for the detection of aneuploidies and gene amplifications by Chromogenic in situ Hybridization (CISH) in formalinfixed, paraffin-embedded tissue sections, cell samples, blood or bone marrow smears, and metaphase chromosome spreads.
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Method Description
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The ZytoDot ® System uses Digoxigenin-labelled probes (1) which are, after blocking (2), detected using a Mouse-anti-Digoxigenin antibody (3). This antibody is detected by a polymerized HRP-Goat-anti-Mouse antibody (4). The enzymatic reaction of DAB (5) leads to the formation of strong permanent brown signals that can be visualized by light microscopy using a 40x objective.
Detection of HER2 gene amplification using the ZytoDot ® SPEC HER2 Probe Kit
No amplification as indicated by 2 signals/dots per nucleus (left). High amplification as indicated by multiple dots and small clusters (middle) or large signal clusters (right).
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Method Description – ZytoDot ® 2C
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The ZytoDot ® 2C System uses Digoxigenin- and DNP-labeled probe cocktails targeting different genomic sections (1). After blocking (2), the probes are detected by using a Mouse-anti-Digoxigenin/Rabbit-anti-DNP antibody-cocktail (3). These antibodies are detected by polymerized HRP-Goat-anti-Mouse (4) and Ap-Goat-anti-Rabbit (5) antibodies. The enzymatic reaction of AP-red (6) and HRP-green (7) leads to the formation of strong permanent red respectively green signals that can be visualized by light microscopy using a 40x objective.
Detection of HER2 gene amplification using the ZytoDot ® 2C SPEC HER2/CEN 17 Probe Kit.
No amplification as indicated by two green (HER2) and two red (CEN 17) signals per nucleus (left). Amplification and chromosome 17 aneusomie as indicated by multiple green and red signals (middle). Strong HER2 amplification with large green clusters (right).
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CISH: A viable alternative to FISH
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High concordance between CISH and FISH ranging from 92-100% has
been shown by numerous international studies for HER2 amplification.
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Advantages of CISH over FISH
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• Quick and easy interpretation of results comparable to IHC
• Simultaneous observation of tissue morphology and CISH signals
• Storage of slides at room temperature – CISH signals are permanent
• No costly fluorescent microscope needed
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Advantages of ZytoDot ® 2C
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• Two targets detected simultaneously
• High contrasting distinct red and green signals
• Visualization at 40x using light microscopy
• Standardized and complete kit solutions
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High Signal-to-Noise Ratio
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All ZytoDot ® probes are processed by the unique ZytoVision ® Repeat Subtraction Technique resulting in advanced specificity and less background. No further blocking of repetitive sequences is needed!
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