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What is In Situ Hybridization?

About this technology

What is In Situ Hybridization?

In situ Hybridization (ISH) is a powerful technique for precise detection and localization of a specific nucleic acid sequence within a histologic section. The underlying principle of ISH is that the nucleic acids within a histologic specimen can be detected by the hybridization of a complementary nucleic acid probe in which a reporter molecule is attached. Common non-radioactive labels for probe are fluorescein and digoxigenin. Subsequent visualization of the reporter molecule localizes DNA or RNA sequences in tissue or cells samples.

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ISH Detection Kits

BioGenex Super Sensitive™ 1-Step Polymer-HRP ISH Detection Kit is suited especially for chromogenic in situ hybridization (CISH) assays. It uses a non-streptavidin-biotin proprietary micropolymer-complex technology to minimize background staining while providing superb probe detection. In situ Hybridization (ISH) is a powerful technique to detect and localize specific nucleic acid sequences within cells or tissues. The hybridization of labeled probes to specific DNA/RNA sequences within a cell allows quantitative detection of that particular target gene. There are two basic approaches to visualizing the DNA or RNA target sequences in situ: CISH and fluorescence in situ hybridization (FISH). BioGenex Super Sensitive™ 1-Step Polymer-HRP ISH Detection Kit was designed especially for CISH

Features and Benefits

  • Highly Sensitive and specific for chromogenic RNA/DNA detection
  • Visualize single copy gene
  • Can be used with third party probes
  • Clean, Intense and reproducible staining.

ISH Probes

BioGenex offers fluorescein-labeled oligonucleotide probes to detect DNA or RNA using In situ Hybridization (ISH). These probes allow quantitative detection of specific DNA/RNA sequences in their native form within the cells of formalin-fixed paraffin-embedded (FFPE) tissue sections. These probes offer reliable, highly sensitive, and easy-to-perform DNA and RNA ISH assays when used with the BioGenex ISH Detection Systems.

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ISH Ancillaries

Ancillaries as a one-stop-shop provider of molecular pathology workstations and reagents, we also offer a comprehensive suite of ancillary products to support our customers. These high-quality products are aimed at accelerating the generation of reproducible and reliable tissue staining results. The ancillaries and consumables, we offer include Antigen Retrievals, Enzymes for Tissue Digestion, Dewaxing and Blocking reagents, Buffers and Diluents, Counter Stains and Mounting media, Vials, Pipette tips, Slides, Coverslips, Pap pen.

Nucleic Acid Retrieval is an effective way to unmask DNA in formalin-fixed, paraffin-embedded (FFPE) tissue sections. The fixation-induced cross-linking of DNA and proteins hampers the hybridization of DNA probes. Using Proteinase K or protease digestion cannot fully dissociate the cross-linking or provide access to the target DNA. Our Nucleic Acid Retrieval solution combined with microwave heating dissociates formalin-induced cross-linking and facilitates better accessibility of DNA to its probes, and improves probe penetration for DNA binding. Therefore, Nucleic Acid Retrieval (NAR-1) is recommended over Proteinase K while using DNA targeting probes.


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  • Peroxide Block: Endogenous peroxidase is most commonly encountered in red blood cells, kidney, and liver tissue. Peroxide Block should be used prior to application of primary antibody when Horseradish Peroxidase (HRP) is used as the labeling enzyme, and if it is necessary to block endogenous peroxidase activity in the tissue being stained.
  • Power Block™: This is a blocking reagent for reducing nonspecific background in immunoassays. A truly universal block, it is suitable for use in immunohistochemistry, immunocytochemistry, ELISA methods, and immunogold techniques. The Power Block reagent contains buffer, casein and preservative and also works well as an antibody diluent and washing medium.
  • Enzymes: Immunoreactivity of an epitope can be recovered using by proteolytic-induced epitope retrieval (PIER). PIER is an enzymatic method of antigen retrieval which relies on enzymes such as proteinase K, and pepsin to unmask the antigenic epitopes by cleaving peptides.


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BioGenex offers the following counterstains for use in Immunohistochemistry, in situ Hybridization and other applications with either manual or automated staining systems.


Mayer’s hematoxylin is a blue stain that does not contain alcohol and therefore is compatible with both alcohol soluble non-permanent chromogens (AEC, Fast Red and New Fuchsin) and alcohol-insoluble chromogens (DAB and Elegance Red). It is alcohol and xylene insoluble and therefore compatible with most clearing agents and mounting media.


Mounting of all stained biological specimens is an essential step before their microscopic evaluation. Mounting also enables the slides to be archived for long periods of time. The mounting medium may be used to attach a coverslip or may itself serve as a coverslip substitute. The choice of mounting medium depends on whether long-term or short-term preservation is desired, and whether the mounting procedure is chemically compatible with the chromogen and the counterstain.

  • SuperMount® Permanent Mounting Medium is a polymer based aqueous mounting media that does not require the use of a coverslip. This innovative, patented mounting medium (BioGenex’s U.S. Patent No. 5,492,837) is designed to preserve biological specimens for long-term storage. SuperMount® medium is compatible with most aqueous and organic-soluble dyes and chromogens including AEC, DAB, Elegance Red, Fast Red, New Fuchsin, BCIP/NBT, Rhodamine, Fluorescein, Texas Red, Phycoerythrin, Phycocyanin, and Fat Stain (Oil Red O). The refractive index of SuperMount® yields greater transparency and clarity of specimens to be examined under the microscope. SuperMount® can be used for the mounting of all biological specimens, including stained tissue sections, Cytospin preparations, and blood smears.
  • Aqueous Mounting Medium is glycerol-based mounting medium that require the use of a coverslip. It is intended for short-term specimen storage and is compatible with most chromogens and counterstains.
  • XMount™ Mounting Medium is a permanent mounting medium that has been optimized for use with BioGenex Xmatrx® instrument for all BioGenex detection systems for immunohistochemistry (IHC), In Situ Hybridization (ISH) and special stains. XMount™ is intended for use with alcohol and xylene insoluble chromogens, such as DAB (for peroxidase systems) and Elegance Red (for alkaline phosphatase systems). XMount dries clear with an ideal refractive index similar to high quality glass and tissue elements. Mounted slides can be viewed with high magnification oil immersion lenses. Also, when mounting preparations stained with the BCIP/NBT substrate, crystal formation that may occur when using other media is minimized.


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BioGenex provides high-quality products which are aimed at accelerating the generation of reproducible and reliable tissue staining results. The consumables, we offer include Vials, Pipette tips, Slides, Coverslips, Pap pen.


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