Fluorescent in situ hybridization (FISH)
Fluorescent in situ hybridization is an important method for genetic mapping and gene expression profiling. This technique exploits the use of fluorescent dye-labeled probes against a nucleotide sequence of DNA that is mapped or localized to a chromosome or intra-nuclear DNA to generate a fluorescence signal which in turn is visualized in situ by microscopy. The hybridization of the probe to the corresponding nucleotide sequence enables detection of the presence or absence of specific sequences, mapping of translocation break points, and determination of gene copy numbers.
As a diagnostic tool, FISH is widely used in tumor diagnosis, clinical genetics, neuroscience, reproductive medicine, toxicology, microbiology, evolutionary biology, comparative genomics, cellular genomics and chromosome biology. It assists in identification of genetic abnormalities associated with prenatal diagnosis, cancer and mental retardation. It is possible to differentiate benign and malignant tumors in the same tissue using FISH. Disruption of regulatory elements in tissues is one of the main causes of tumor susceptibility which has been exploited to understand the aberrant gene functions. The traditional methods of morphological diagnosis are restricted in their use because tumors appear similar under a microscope. However, FISH overcomes this ambiguity by characterizing the genetic mutations as biomarkers.
FISH is also an important tool for pathologists to diagnose infectious disease and cancer. It also helps make treatment decisions. For example, BRCA1/2 status is an important cancer marker that aid treatment decisions and can be detected by FISH.
Automated FISH diagnostic systems have been approved by the US FDA under the medical device stream, a mark of the maturity and reliability of the technique.
BioGenex eFISH Solution:
BioGenex eFISH system consist of high specificity eFISH probes, visualization kits, ancillary reagents, in addition to our brand new automated systems: NanoVIP®, Xmatrx (Elite & Infinity) Xmatrx® NANO, and the Xmatrx® MINI. These systems are open adaptable to your laboratories workflow. The BioGenex eFISH probes provide improved signal intensity. This results from proprietary design by Repeat Subtraction Technique. Our eFISH probes and visualization kits seamlessly integrate with our full automation, semi automation or manual system and provide reliable, sharp, and bright signals.
BioGenex offers eFISH probes for oncology, and hematology to include dual and triple color probes, break apart, fusion, and enumeration probes. eFISH Histo (formalin-fixed paraffin-embedded sections) and Cyto (cytology Specimen) Visualization Kits are standardized with ready-to-use reagents including protease and buffer for pre-treatment and post hybridization stringency washes. Kits include template protocols optimized to provide reliable and reproducible intense stains with minimum background thus avoiding repeats.
In the eFISH procedure, fixed tissue sections are pretreated to expose target DNA or miRNA sequences. An appropriately labelled probe is hybridized to the exposed target DNA or miRNA sequences in the cells. Subsequent stringent washing steps remove any probe that is non-specifically bound to the tissue section. Subsequently slides are mounted using DAPI/antifade and can be visualized under a fluorescence microscope using the appropriate filter set. BioGenex provides a wide range of eFISH probes as well as automated systems.
Nucleic Acid Retrieval is an effective way of unmasking DNA in formalin-fixed, paraffin-embedded tissue sections using microwave heating. BioGenex’s Nucleic Acid Retrieval solution breaks the formalin-induced cross-linking bonds between DNA and proteins, as well as protein-protein cross-linking thereby allowing better penetration of probes and accessibility of DNA for binding. Nucleic Acid Retrieval (NAR-1) is recommended instead of Proteinase K when DNA targeting probes are used.
BioGenex eFISH Visualization kit is intended for use in a Fluorescence in situ Hybridization (FISH) procedure and is optimized for the detection of target nucleic acid sequence. It is designed for the specific fluorescence detection of fluorescent-labelled nucleic acid probes following hybridization to target DNA or miRNA sequences.
eFISH Visualization kits are available for formalin-fixed, paraffin-embedded (FFPE) tissue sections and cell preparations. BioGenex eFISH visualization kits provide unsurpassed performance.
The BioGenex eFISHHisto kit is a novel system for processing formalin-fixed, paraffin-embedded (FFPE) tissues. This kit contains reagents for pretreatment and post-hybridization stringency washes of the tissue. This kit is to be used for BioGenex eFISHiency FISH processing platforms Xmatrx® Infinity/ELITE, Xmatrx® NANO, NanoVIP® and Xmatrx® MINI along with BioGenex eFISH probes.
The BioGenex eFISH cytology kit is a novel system for processing of cytological/hematological specimens/smears/cells. This kit contains reagents for pretreatment and post-hybridization stringency washes of the tissue. This kit is to be used for BioGenex eFISHciency FISH processing platforms Xmatrx® Infinity/Elite, Xmatrx® NANO, NanoVIP® and Xmatrx® MINI along with BioGenex eFISH probes.
DAPI Mounting Medium contains 4’, 6-diamidino-2-phenylindole (DAPI), which fluoresces when bound to DNA and is used as a nuclear or chromosomal counterstain. DAPI produces a blue fluorescence when bound to DNA with excitation at about 360nm and emission at 460nm. DAPI Mounting Medium is a unique, stable formula for preventing photo-bleaching of fluorescent proteins and fluorescent dyes during microscopic examination of fluorescence in situ hybridization specimens. DAPI Mounting Medium is water-soluble and has been optimized to preserve fluorescence during long-term storage, if mounted slides are stored at 2-8°C.
DAPI Mounting Medium does not solidify, but remains as liquid on the slide and can be stored without sealing. To seal the coverslips permanently onto the slides, apply nail polish or a plastic sealant around the perimeter. Mounted slides should be stored at -20°C protected from light. After use, store the remaining DAPI Mounting Medium at -20°C and protected from light.
As a one-stop shop provider of molecular pathology workstations and reagents, BioGenex offer a comprehensive suite of ancillary products to support our customers including enzymes for tissue processing, dewaxing and rehydrating reagents, buffers, microscope slides and coverslips, and pipette tips. These high-quality products are aimed at accelerating the generation of reproducible and reliable tissue staining results.