ZytoVision

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ZytoLight ® products are designed for the identification of genetic aberrations e.g. translocations, deletions, amplifications, and chromosomal aneuploidies by Fluorescence in situ Hybridization (FISH) in formalin-fixed, paraffin-embedded tissue sections, cell samples, blood or bone marrow smears, and metaphase chromosome spreads.

Method Discription

The ZytoLight system uses direct labeled FISH probes (1), eliminating the need to detect the probes with fluorophore-coupled antibodies. The probes are detected by fluorescence microscopy using appropriate filter sets (2). Due to an exciter filter (3), full-spectrum light, emitted by the microscope lamp (4), is reduced to light of a defined wavelength that specifically excites the fluorophore of the probe. This light is reflected onto the specimen by a dichroic mirror (5). The fluorophore emits light of longer wavelengths that passes the mirror. Finally, a barrier filter (6) reduces the emitted light to a defined wavelength that can be detected.

ZytoLight ® Fluorochromes

Two factors that mainly influence FISH analyses:
  • Fluorochromes of the FISH probes
  • Appropriate filter sets

The ZytoDot ® and the ZytoDot ® 2C products are designed for the detection of aneuploidies, gene amplifications or gene deletions and translocations by Chromogenic in situ Hybridization (CISH) in formalin-fixed, paraffin-embedded tissue sections, cell samples, blood or bone marrow smears, and metaphase chromosome spreads.

Method Description – ZytoDot ®

Single Color CISH for the Detection of Genomic Alterations

The ZytoDot system uses Digoxigenin-labeled 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.

Method Description – ZytoDot ® 2C

2-Color CISH for the Detection of Genomic Alterations

The ZytoDot 2C system uses DIG- and DNP-labeled probe cocktails targeting different genomic sections (1) which are detected using a Mouse-anti-DIG/Rabbit-anti-DNP cocktail (2). These antibodies are detected by a unique cocktail of polymerized HRP-Goat-anti-Mouse/AP-Goat-anti-Rabbit antibodies (3). The enzymatic reaction of AP-Red (4) and HRP-Green (5) leads to the formation of strong permanent red respectively green signals that can be visualized by light microscopy using a 40x objective.

Advantages of CISH

  • Simultaneous observation of tissue morphology and CISH signals
  • Quick and easy interpretation of results comparable to IHC
  • Storage of slides at room temperature - CISH signals are permanent
  • No costly fluorescent microscope needed

The ZytoFast ® products are designed for outstandingly fast detection and discrimination of human pathogen viruses, e.g. HPV, EBV, CMV, and the determination of lymphocyte clonality by detecting Ig-κ and Ig-λ light chain RNA by Chromogenic in situ Hybridization (CISH) in formalin-fixed, paraffin-embedded tissue sections and cell samples. The signal intensity of ZytoFast probes is increased even more when using the ZytoFast PLUS Implementation Kits.

Method Description – ZytoFast ®

The ZytoFast system uses oligonucleotide probes tagged with Biotin (1) which are detected using enzyme-conjugated streptavidin targeting the tag (2).
The enzymatic reaction of chromogenic substrates (3), e.g. BCIP/NBT or AEC, leads to the formation of strong color precipitates that can be visualized by light microscopy.

Method Description – ZytoFast ® PLUS

The ZytoFast PLUS system uses Digoxigenin-labeled probes (1) which are detected using primary antibodies (2). These antibodies are detected by polymerized enzyme-conjugated secondary antibodies (3). The enzymatic reaction of chromogenic substrates (4), e.g. NBT/BCIP, AEC or DAB, leads to the formation of strong color precipitates that can be visualized by light microscopy.

Advantages of Fast-CISH

  • Simultaneous observation of tissue morphology and CISH signals
  • No risk of false positives due to mispriming or contamination as with PCR
  • Easy method comparable to IHC
  • No costly equipment needed
  • Ability to test archival specimens
  • High sensitivity and specificity