Remaining IHC SOPs
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Questions and Answers

What is the purpose of the positive control when performing IHC?

  • To assess the presence of specific immunohistochemistry staining
  • To assess the presence of non-specific staining (correct)
  • To assess the performance of the primary antibody
  • To verify and accept all controls and test slides

    • True or false: Immunohistochemistry (IHC) is used to identify proteins in tissue.

    True

    The purpose of the negative control is to assess the presence of ______ staining.

    specific immunohistochemistry

    Immunohistochemistry (IHC) is used to identify the presence of ______ in tissue.

    <p>proteins</p> Signup and view all the answers

    True or false: Positive controls are used to assess the presence of non-specific staining.

    <p>True</p> Signup and view all the answers

    How is the Ventana Benchmark Ultra software used?

    <p>To register the dispenser and the reagent</p> Signup and view all the answers

    The purpose of the positive control is to assess the presence of ______ staining.

    <p>non-specific</p> Signup and view all the answers

    Quality control for IHC is performed and documented according to ______ and manufacturer recommendations.

    <p>specific standard operating procedures</p> Signup and view all the answers

    True or false: Negative controls are used to assess the presence of specific immunohistochemistry staining.

    <p>True</p> Signup and view all the answers

    What is the purpose of the ISH iVIEWBlue Plus Detection Kit?

    <p>To detect specific DNP labeled probes and antibodies bound to a target sequence or antigen in paraffin-embedded tissue sections</p> Signup and view all the answers

    The procedure for verifying and accepting new reagent lots is ______.

    <p>described</p> Signup and view all the answers

    The purpose of the negative control is to assess the presence of ______ staining.

    <p>specific immunohistochemistry</p> Signup and view all the answers

    What is the purpose of parallel staining?

    <p>To control for variables such as disparity in the lots of detection reagents or instrument function</p> Signup and view all the answers

    True or false: Quality control for IHC is performed according to manufacturer recommendations.

    <p>True</p> Signup and view all the answers

    The Ventana Benchmark Ultra software will be used to ______ the dispenser and the reagent has been received and needs to be QCd.

    <p>register</p> Signup and view all the answers

    Quality control for IHC is performed and documented according to ______ and manufacturer recommendations.

    <p>specific standard operating procedures</p> Signup and view all the answers

    The IHC technician will enter concentration and expiration within the same ______, highlight the prep kit number and click on fill using the correct ml, then close.

    <p>window</p> Signup and view all the answers

    True or false: The ISH iVIEWBlue Plus Detection Kit detects specific DNP labeled probes and antibodies in paraffin-embedded tissue sections.

    <p>True</p> Signup and view all the answers

    The IHC technician will place small syringe containing ______ into the top of the dispenser.

    <p>residual reagent</p> Signup and view all the answers

    What is the purpose of quality control for IHC?

    <p>To perform according to specific standard operating procedures and manufacturer recommendations</p> Signup and view all the answers

    The prepared slides will be documented on the ______ and given to the pathologist for signature.

    <p>New Reagent Lot of Acceptability Form</p> Signup and view all the answers

    True or false: The Ventana Benchmark Ultra software is used to register the dispenser and the reagent.

    <p>True</p> Signup and view all the answers

    How is the reagent transferred into the dispenser?

    <p>By opening the cap on the top of the dispenser and pouring the reagent into a large syringe</p> Signup and view all the answers

    The Ventana Benchmark Ultra software will be used to register the ______.

    <p>dispenser</p> Signup and view all the answers

    True or false: The prepared slides need to be documented on the New Reagent Lot of Acceptability Form.

    <p>True</p> Signup and view all the answers

    The ISH iVIEWBlue Plus Detection Kit detects ______ and antibodies bound to a target sequence or antigen in paraffin-embedded tissue sections.

    <p>specific DNP labeled probes</p> Signup and view all the answers

    The ISH iVIEWBlue Plus Detection Kit detects specific DNP labeled ______ and antibodies.

    <p>probes</p> Signup and view all the answers

    What is the purpose of the negative control when performing IHC?

    <p>To assess the presence of specific immunohistochemistry staining</p> Signup and view all the answers

    What is the purpose of the enzyme-labeled secondary antibody or biotin-conjugated secondary antibody used in the ISH iVIEWBlue Plus Detection Kit?

    <p>To localize the labeled probe or antibody</p> Signup and view all the answers

    The complex is then visualized with ______ and nitro blue tetrazolium (NBT) chromogen.

    <p>5-Bromo-4-chloro-3-indolyl phosphate (BCIP)</p> Signup and view all the answers

    True or false: The complex is visualized with 5-Bromo-4-chloro-3-indolyl phosphate (BCIP) and nitro blue tetrazolium (NBT) chromogen.

    <p>True</p> Signup and view all the answers

    This step is followed by the addition of a ______ enzyme conjugate which binds to the biotin present on the secondary antibody.

    <p>Streptavidin-AP</p> Signup and view all the answers

    Parallel staining is required to control for ______.

    <p>variables</p> Signup and view all the answers

    What is the difference between a "true positive" stain and a "false positive" stain?

    <p>A true positive stain shows chromogen deposition in cells or structures that truly contain the antigen of interest, while a false positive stain is one where the chromogen is localized to cells or structures that in reality lack the antigen of interest.</p> Signup and view all the answers

    A "true positive" stain shows chromogen deposition in cells or structures that ______ contain the antigen of interest.

    <p>truly</p> Signup and view all the answers

    True or false: A "false positive" stain is one where the chromogen is localized to cells or structures that in reality contain the antigen of interest.

    <p>False</p> Signup and view all the answers

    Parallel staining is required to control for variables such as ______ in the lots of detection reagents or instrument function.

    <p>disparity</p> Signup and view all the answers

    What is the purpose of the New Reagent Lot of Acceptability Form?

    <p>To document confirmation of acceptance of new reagent lot results</p> Signup and view all the answers

    When slides are returned to the Immunohistochemistry (IHC) section, document ______ of acceptability in the antibody data sheet binder.

    <p>confirmation</p> Signup and view all the answers

    True or false: Parallel staining is required to control for instrument function.

    <p>True</p> Signup and view all the answers

    Study Notes

    • Immunohistochemistry (IHC) is a technique used to identify the presence of specific proteins in tissue.
    • Positive and negative controls are used to determine the performance of the primary antibody.
    • The purpose of the positive control is to assess the presence of non-specific staining.
    • The purpose of the negative control is to assess the presence of specific immunohistochemistry staining.
    • A pathologist must verify and accept all controls and test slides for IHC.
    • Quality control for IHC is performed and documented according to specific standard operating procedures and manufacturer recommendations.
    • The procedure for performing lot-to-lot testing is described.
    • The procedure for verifying and accepting new reagent lots is described.
    • The procedure for making the IHC test labels is described.
    • The IHC technician will open the cap on the top of the dispenser while it is standing in an upright position, pour reagent into a large syringe, replace the syringe plunger, and place the filter on the end of the large syringe. They will then depress the large syringe to transfer the reagent into the dispenser.
    • The IHC technician will then place small syringe containing residual reagent into the top of the dispenser and then depress the small syringe, transferring the reagent back into the dispenser.
    • The Ventana Benchmark Ultra software will be used to register the dispenser and the reagent has been received and needs to be QCd.
    • The IHC technician will enter concentration and expiration within the same window, highlight the prep kit number and click on fill using the correct ml, then close. The prepared slides will be documented on the New Reagent Lot of Acceptability Form and given to the pathologist for signature.
    • The ISH iVIEWBlue Plus Detection Kit detects specific DNP labeled probes and antibodies bound to a target sequence or antigen in paraffin-embedded tissue sections.
    • The labeled probe or antibody is located by an anti-DNP antibody, then by an enzyme-labeled secondary antibody or a biotin-conjugated secondary antibody.
    • This step is followed by the addition of a Streptavidin-AP (alkaline phosphatase) enzyme conjugate which binds to the biotin present on the secondary antibody.
    • The complex is then visualized with 5-Bromo-4-chloro-3-indolyl phosphate (BCIP) and nitro blue tetrazolium (NBT) chromogen, which produces a blue precipitate that is readily detected by light microscopy.
    • A "true positive" stain shows chromogen deposition in cells or structures that truly contain the antigen of interest. In contrast, a "false positive" stain is one where the chromogen is localized to cells or structures that in reality lack the antigen of interest.
    • The purpose of this procedure is to compare new antibody/probe detection system reagent lot numbers to previous lot numbers concurrently or before being placed into service.
    • For Immunohistochemistry (IHC) parallel staining using one known positive and one known negative will be performed on the same instrument with the two detection kits using the different multi-tissue control tissue.
    • Parallel staining is required to control for variables such as disparity in the lots of detection reagents or instrument function.
    • The IHC staining has to be performed with the old lot on the first run and the new lot on the same run (utilizing the landing zone feature on the Ventana software for the new lot) due to the fact the instrument will select the older reagent if both lots were placed at the beginning of the run.
    • After staining is complete, the slides are given to the designated pathologist for review. When slides are returned to the Immunohistochemistry (IHC) section, document confirmation of acceptance of new reagent lot results on the IHC detection system comparison form for detection and document antibody confirmation of acceptability in the antibody data sheet binder.

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