Buffers and Diluents
Buffers and diluents are available for Immunohistochemistry, in situ Hybridization Special Stains and most other applications.
- General buffers, such as PBS (PH 7.6) and TBS (PH 7.6, 0.1M) can be used for washing/rinsing of slides.
- Super SensitiveTM Wash Buffer is a phosphate buffered saline (PH 7.4) with surfactant and is used to ensure optimal staining with even spreading of antibodies and other reagents to avoid inconsistent results.
- Common Antibody Diluent and Enhanced Antibody Diluent have been developed for use with all antibodies in immunohistochemistry and have been specifically optimized for use with BioGenex antibodies and reagent products. These diluents enable enhancement of signal-to-noise ratio of staining when used optimally and also help in maintaining the antibody specificity and stability. The Enhanced Antibody Diluent, in addition to all the above features, contains chemical compounds to enhance antigen-antibody interaction and affinity. In order to achieve desired staining pattern and intensity, the titers of antibodies or concentrations of reagents may need to be optimized. These diluents are also for diluting concentrated Alkaline Phsophatase (AP) labels but are not suitable for diluting Horseradish Peroxidase (HRP) labeles because they contain Sodium Azide (0.09%).
- Streptavidin Peroxidase Diluent, was developed especially for diluting concentrated HRP labeles and does not contain Sodium Azide.
- Link Diluent, was developed for diluting concentrated Link (Biotinylated Anti-Immunoglobulins) antibodies.
- 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 BlockTM: 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.
- Protein Block: This can be used to reduce background staining due to non-specific binding of the primary or secondary antibodies to the tissue. Protein Block should be applied immediately prior to the primary antibody.
Counterstains and Mounting Media
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.
- DAPI (4',6-diamidino-2-phenylindole) is a fluorescent blue stain used as a nuclear counterstain in Fluorescent In Situ Hybridization (FISH) and Immunofluorescence (IF) applications. DAPI strongly binds A-T rich regions in DNA and can be used to stain nuclei of both live and fixed cells. DAPI has an absorption maximum at a wavelength of 358 nm and its emission maximum is at 461 nm.
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.
- XMountTM Mounting Medium is a permanent mounting medium that has been optimized for use with BioGenex XMount 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.
The chart below summarizes the compatibility of mounting medium, chromogens and counterstains.
|Chromogen||Stain Color||Enzyme used||Solubility in Alcohol/Xylene||Compatible with Hematoxylin||Compatible Mounting Media|
|AEC||Brick Red||HRP||Yes||Yes||Aqueous or Super Mount|
|DAB||Brown||HRP||No||Yes||Aqueous, Super Mount or XMount|
|Elegance Red||Red||AP||No||Yes||Aqueous, Super Mount or XMount|
|Fast Red||Red||AP||Yes||Yes||Aqueous or Super Mount|
|New Fuchsin||Red||AP||Yes||Yes||Aqueous or Super Mount|
Reagent Vials & Accessories
1. i6000 Elite & Xmatrx Infinity: The OptiMiser reagent vials (U.S. & Foreign Equivalent Patents Pending) are available as a 20 ml disposable pack for use on the i6000TM or Xmatrx Infinity staining systems. Vials are designed to minimize dead volume:
|XT026-V24||Small White vials (20 ml), including vial holders XT027-H24.
(24 per pack)
|XT101-24X||Small White vials (20 ml), including vial holders XT027-H24.
(24 per pack)
|XT126-24V||Brown vial without Neck/lid as holder for 2 ml insert, , including vial holders XT027-H24.
(24 per pack)
|XT149-V24||Vial Insert – 2 ml
(24 per pack)
|XT027-H24||Vial holders for 20 ml vials
(24 per pack)
2. Xmatrx Elite:: Reagents vials for Xmatrx Elite Automated Staining Systems are RFID-tagged 17 ml vials especially designed to ensure accurate identification, proper reagent inventory management and staining of up to 200 slides. These vials dead volume is minimized to <0.5 ml. RFID-tagged vials for use with antibodies from any supplier (user defined) are also available (XT077-AX601 to XT077-AX0999).
ISH probes are supplied in 2 ml vials, inserted in RFID-tagged vial holders thus minimizing dead volume to <0.05 ml. RFID-tagged vials for use with ISH probes from any supplier (user defined) are also available (XT079-PR0050 to XT079-PR0099).
Microscope Slides & Coverslips
OptiPlusTM Positive-Charged Microscope Slides provide a strong adhesive surface for tissues and cells to prevent tissue displacement during harsh pre-treatments such as enzymatic digestion and the microwave Antigen Retrieval method. These slides are ideal for automated systems. Additionally, each slide has a frosted end for easy labeling. The OptiPlus Positive-Charged Barrier Slides have all the advantages of our regular OptiPlus slides, but also contain hydrophobic barriers that allow the quantity of reagents per slide to be tailored to the size of the specimen. These slides eliminate reagent waste without the need to use a PAP pen, thereby reducing set-up time in manual assays as well as in automated systems. The permanent hydrophobic barriers are compatible with dewaxing solutions and other reagents. The slides are suitable for use with frozen tissue sections, formalin-fixed paraffin sections, and cytology preparations.
1. i6000 Elite Automated Staining Systems:
OptiPlusTM Barrier slides for i6000 come in three different configurations to accommodate different tissue sizes or multiple tissues per slide:
- A single, full-size test area of 25 x 40 mm
- A single 2/3-size test area of 25 x 30 mm
- Three 1/3-size test areas per slide, each measuring 25 x 15 mm
2. Xmatrx Automated Staining Systems:
OptiPlusTM Barrier Slides for Xmatrx (U.S. & Foreign Equivalent Patents Pending) contain a double hydrophobic barrier that allows formation of an oil seal to prevent evaporation of micro-reagents during high temperature steps and prolonged incubations. Four different configurations are available:
- A single test area of 25 x 40 mm (>80 µl of reagent recommended)
- A single test area of 25 x 25 mm (>40 µl of reagent recommended)
- A single test area of 18 x 18 mm (>10 µl of reagent recommended)
- Two test area per slide, each measuring 18 x 18 mm
Coverslips are optimized for use on Xmatrx staining systems and come is three configurations to accommodate the different barrier slides.
BioGenex pipette tips are made of high-quality polypropylene and are RNase and heavy metal-free when untampered. Inner surface is extremely smooth and requires minimum wetting. 1 ml pipette tips are optimized for use on BioGenex Xmatrx and i6000 Staining Systems, while 200 µl tips are optimized for Xmatrx staining systems.
Consumables kits for Xmatrx
|Item||SKU||Size||Barrier Slides 25x40 mm||Barrier Slides 25x25 mm||Coverslips 25x40 mm||Coverslips 25x25 mm||1 ml Pipette Tips||200 ml Pipette Tips|
|IHC kit||XT148-YCD||200 test||216||NA||1000||NA||384||960|
|ISH kit||XT144-YAD||100 test||NA||104||NA||900||384||960|
1. Antigen Retrieval Accessories Kits
The Antigen Retrieval Accessory Kit consists of slide holders and slide baths that make it convenient and compatible with any of the several Antigen Retrieval solutions. To accommodate microwave heating, the slide baths and slide holders are made of heat-stable thermoplastic polyolefin and hydrocarbon polymers of acetal resins. These accessories may be used in a microwave or a pressure cooker.
|Item||SKU||Slide Bath + Lid||Slide Holder|
|24-Slide Accessory kit||MW001-SU||1||1(24-slide capacity)|
|72-Slide Accessory kit||MW001-HB||3||3(72-slide capacity)|
2. NordicWare® Microwave Pressure Cooker
Placing the NordicWare Microwave Pressure Cooker within a microwave is an effective method for enhancing staining with the Antigen Retrieval technique. The heat produced under enhanced pressure can reduce the build up of gas bubbles on the surface of tissues. This improves the intensity of staining, accompanied by preservation of tissue and cell morphology. This pressure cooker is also optimized for use with various BioGenex Antigen Retrieval solutions. BioGenex Catalog number: NW001-PC.
3. PAP Pen for Tissue Staining
The PAP Pen is a useful pen-like tool for immunohistochemical staining methods. It is designed to prevent the waste of valuable reagents by forming a water-repellent barrier around the specimen. This barrier creates the proper surface tension to hold an antibody solution or detection reagents within the target area on the slide. The surface tension provided by the PAP Pen circle ensures that only the amount of antibody solution needed for sufficient reaction will be applied. Since over-flooding of the slide is eliminated, wiping of excess fluid around the specimen can be avoided. The PAP Pen can be used for immunostaining of paraffin sections, frozen sections, and for fluorescent antibody methods. The PAP Pen contains a special formulation, which is water repellent. It can be removed, if desired, with xylene or xylene substitutes after the staining procedure is completed. BioGenex Catalog Number: XT001-PP, sufficient for use on 500-1000 slides.