Immunohistochemistry Procedure: Paraffin Embedded Tissue Processing
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Questions and Answers

What is the purpose of blocking nonspecific sites in immunohistochemistry?

  • To enhance the signal of the antigen-antibody complex
  • To minimize background staining (correct)
  • To activate endogenous enzyme
  • To prevent the primary antibody from binding to the epitope

    • What is the main purpose of antigen retrieval in immunohistochemistry?

  • To conjugate the primary antibody
  • To visualize the antigen-antibody complex
  • To enhance the binding of the secondary antibody
  • To unmask epitopes masked by fixation (correct)

    • What is the role of the secondary antibody in immunohistochemistry?

  • To activate endogenous enzyme
  • To bind to the primary antibody (correct)
  • To recognize the epitope on the antigen
  • To enhance the signal of the antigen-antibody complex

    • What is the purpose of dehydration and clearing in immunohistochemistry?

    <p>To prepare the tissue for paraffin embedding</p> Signup and view all the answers

    What is the epitope in immunohistochemistry?

    <p>The region on the antigen recognized by the antibody</p> Signup and view all the answers

    What is the primary function of epitope retrieval?

    <p>To break protein cross-links formed by fixation</p> Signup and view all the answers

    What is the purpose of proteolytic induced epitope retrieval (PIER)?

    <p>To selectively break protein links to reveal hidden epitopes</p> Signup and view all the answers

    What is the mechanism by which heat induced epitope retrieval (HIER) works?

    <p>By using specific buffered solutions to recover antigen form</p> Signup and view all the answers

    What is the term for the binding site on an antibody that recognizes an epitope?

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

    What is the result of cross-linking of amino acids within an epitope?

    <p>Masking of epitopes</p> Signup and view all the answers

    What is the term for the protein produced by B lymphocytes in response to the presence of an antigen?

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

    What is the region on an antigen that a specific antibody binds to?

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

    What is the result of injecting an antigen into a rabbit?

    <p>Activation of B cells</p> Signup and view all the answers

    What type of antibody is produced by a single antibody-producing B cell?

    <p>Monoclonal antibody</p> Signup and view all the answers

    What is the function of plasma B cells?

    <p>To produce antibodies</p> Signup and view all the answers

    What is the result of obtaining antiserum from a rabbit?

    <p>Production of polyclonal antibodies</p> Signup and view all the answers

    What is the characteristic of a polyclonal antibody?

    <p>Recognizes multiple epitopes on the same antigen</p> Signup and view all the answers

    What is the primary purpose of blocking nonspecific sites during immunohistochemical staining?

    <p>To increase the specificity of the antibody-antigen interaction</p> Signup and view all the answers

    Which of the following is NOT a step in the processing of paraffin-embedded tissue for immunohistochemical analysis?

    <p>western blotting</p> Signup and view all the answers

    What is the purpose of the biotin/avidin complex (ABC) or labelled streptavidin biotin (LSAB) in indirect immunohistochemical techniques?

    <p>To amplify the signal generated by the primary antibody</p> Signup and view all the answers

    What is the primary advantage of using direct immunohistochemical techniques over indirect techniques?

    <p>Simplified protocol</p> Signup and view all the answers

    What is the purpose of antigen retrieval in immunohistochemical analysis?

    <p>To restore the antigenic sites masked during tissue processing</p> Signup and view all the answers

    What is the primary goal of immunohistochemistry in tumour pathology?

    <p>To define cell types</p> Signup and view all the answers

    Which of the following is a potential application of immunohistochemistry?

    <p>Evaluating immune complex deposition in autoimmune disease</p> Signup and view all the answers

    What is the function of antibodies in immunohistochemistry?

    <p>To detect cellular components in tissue sections</p> Signup and view all the answers

    What is the role of the variable (V) domains in immunohistochemistry?

    <p>Antigen-binding domains</p> Signup and view all the answers

    Which of the following is NOT a application of immunohistochemistry?

    <p>Prenatal diagnosis</p> Signup and view all the answers

    What is the significance of immunohistochemistry in identifying potential target therapy strategies?

    <p>It aids in identifying predictive markers</p> Signup and view all the answers

    Study Notes

    Blocking Nonspecific Sites

    • Prevents non-specific binding of antibodies to the tissue section

    Antigen Retrieval

    • Exposes epitopes masked by formalin fixation for antibody binding

    Secondary Antibody

    • Binds to the primary antibody, enabling signal amplification or visualization

    Dehydration and Clearing

    • Removes water and replaces it with a solvent compatible with embedding medium

    Epitope

    • Specific region on an antigen that an antibody recognizes

    Epitope Retrieval

    • Unmasks hidden epitopes by breaking bonds that keep them inaccessible

    Proteolytic Induced Epitope Retrieval (PIER)

    • Uses enzymes to cleave proteins and expose epitopes

    Heat Induced Epitope Retrieval (HIER)

    • Uses heat to break protein bonds, making epitopes accessible

    Paratope

    • The antigen-binding site on an antibody

    Cross-linking of Amino Acids

    • Altering an epitope's structure and potentially disrupting antibody binding

    Antibody

    • Proteins produced by B lymphocytes in response to antigens

    Epitope

    • Specific region on an antigen that a specific antibody binds to

    Antiserum Production

    • Injecting antigens into a rabbit triggers antibody production

    Monoclonal Antibody

    • Antibody produced by a single antibody-producing B cell

    Plasma B Cells

    • Produce antibodies in response to antigens

    Antiserum

    • Contains antibodies specific to the injected antigen

    Polyclonal Antibody

    • Mixture of antibodies that recognize different epitopes on the same antigen

    Purpose of Blocking

    • Blocks non-specific antibody binding to prevent false positive results

    Immunohistochemistry Processing

    • Not a step in processing: Direct visualization of antigens by antibodies coupled to fluorescent molecules

    Biotin-Avidin Complex (ABC) or Labelled Streptavidin Biotin (LSAB)

    • Amplify the signal for visualization in indirect immunohistochemistry

    Direct Immunohistochemical Techniques

    • Advantage: Simpler, fewer steps, reduced risk of background staining

    Antigen Retrieval Goal

    • Expose epitopes masked by fixation or tissue processing for optimal antibody binding

    Immunohistochemistry in Tumor Pathology

    • Primary goal: To identify and characterize tumor cells

    Applications of Immunohistochemistry

    • Diagnosis, prognosis, and monitoring of disease

    Function of Antibodies

    • Bind to specific antigens and allow their visualization

    Variable (V) Domains

    • Responsible for recognizing specific epitopes on antigens

    Immunohistochemistry - Not an Application

    • Monitoring blood glucose levels (used for diabetes management)

    Immunohistochemistry for Target Therapy

    • Helps identify specific biomarkers on tumor cells, leading to personalized targeted therapies

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    Description

    Test your knowledge of the immunohistochemistry procedure, including paraffin embedded tissue processing, from harvesting and formalin fixation to image capture and analysis. This quiz covers the steps involved in indirect tissue processing, including dehydration, clearing, dissection, embedding, microtomy, and more.

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