H&E Staining Mechanism Quiz lecture 5 除了part2
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Questions and Answers

What is the purpose of using DPX or XPF on coverslip glass in the alternative dry mount method?

  • To increase the visibility of calcium deposits
  • To prevent the staining from fading over time (correct)
  • To improve the differentiation of acid alcohol
  • To enhance the staining of nuclei
  • What is the color of connective tissue as observed in the H&E results?

  • Grey/blue
  • Pale pink (correct)
  • Deep blue
  • Red
  • What is the potential impact of varying the thickness of cut sections on H&E staining variability?

  • Enhances the differentiation using acid alcohol
  • Reduces the staining intensity of muscle fibres
  • Affects the balance of staining time and concentration of the nuclear stain (correct)
  • Improves the visualization of eosinophilic granules
  • What is the recommended thickness for achieving an optimum balance in H&E staining?

    <p>4 micron</p> Signup and view all the answers

    Which of the following is the mechanism of staining used in the direct method?

    <p>Attraction of acidic elements to basic dyes</p> Signup and view all the answers

    What is the function of a mordant in the indirect method of staining?

    <p>To link the tissue and dye</p> Signup and view all the answers

    What is the color of the dye lake produced when a mordant is combined with a dye?

    <p>Dependent on the specific mordant used</p> Signup and view all the answers

    What is the process involved in progressive staining?

    <p>Up to the point for the desired depth of coloration to be achieved</p> Signup and view all the answers

    Which tissue element has negative charges and reacts with haematoxylin?

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

    What reaction occurs when haematoxylin, a basic dye, reacts with acidic nuclei?

    <p>Blue/black coloration</p> Signup and view all the answers

    What color does eosin, an acid dye, produce when it reacts with basic tissue elements?

    <p>Red/shades of pink</p> Signup and view all the answers

    What is the purpose of regressive staining?

    <p>Overstaining of tissue followed by removal of the stain</p> Signup and view all the answers

    What is the charge of RBCs, cytoplasm, muscle, and collagen?

    <p>Basic (+ve charges)</p> Signup and view all the answers

    What reaction occurs when eosin, an acid dye, reacts with RBCs, cytoplasm, muscle, and collagen?

    <p>Red/shades of pink coloration</p> Signup and view all the answers

    What is the function of Haematoxylin in the H&E staining technique?

    <p>To illustrate nuclear detail in cells</p> Signup and view all the answers

    Which substance is oxidized to produce hematein, the actual dye used in an H&E stain?

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

    What is the purpose of using a mordant such as aluminum ammonium sulfate (alum) in the H&E staining process?

    <p>To improve the bonding of hematein to the anionic tissue components</p> Signup and view all the answers

    What is the primary function of Eosin in the H&E staining technique?

    <p>To counterstain and distinguish between the cytoplasm and nuclei of cells</p> Signup and view all the answers

    Why is filtering required for Harris hematoxylin, a strong, regressive stain?

    <p>To remove precipitates</p> Signup and view all the answers

    Which substance is known for fading rapidly in sunlight, requiring labs to use Eosin Y as a counterstain?

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

    What step follows haematoxylin staining in the basic protocol for H&E staining?

    <p>Differentiating with acid alcohol</p> Signup and view all the answers

    What is responsible for staining cytoplasm in H&E staining?

    <p>Harris hematoxylin</p> Signup and view all the answers

    What solution is commonly used for differentiating Harris hematoxylin in most labs?

    <p>1% or 0.5% HCl in 70% alcohol</p> Signup and view all the answers

    What step follows counterstaining with eosin in the staining schedule for H&E?

    <p>Differentiating with acid alcohol</p> Signup and view all the answers

    What feature is very crisp in an example of H&E staining?

    <p>Microvilli on the columnar epithelium</p> Signup and view all the answers

    Study Notes

    • Hematoxylin and Eosin (H&E) is the most commonly used staining technique in histology for diagnosing malignancies.
    • Haematoxylin, derived from Haematoxylon campechianum (bloodwood), is used to illustrate nuclear detail in cells, with the depth of coloration depending on the length of time the sample spends in haematoxylin.
    • Haematoxylin is oxidized to produce hematein, which is the actual dye used in an H&E stain, and the use of a mordant such as aluminum ammonium sulfate (alum) improves the bonding of hematein to the anionic tissue components, most commonly chromatin.
    • Eosin, a red crystalline dye, is the most commonly used counterstain that distinguishes between the cytoplasm and nuclei of cells.
    • Histopathology diagnosis relies heavily on H&E staining, demonstrating a broad range of cytoplasmic, nuclear, and extracellular matrix features, and producing a crisp nuclear staining with Harris hematoxylin.
    • Harris hematoxylin is a strong, regressive stain that produces well-defined nuclear outlines but requires filtering to remove precipitates, and most labs use an aggressive differentiating solution like 1% or 0.5% HCl in 70% alcohol for a few seconds.
    • Eosin is not permanent and fades rapidly in sunlight, with Eosin Y being the most commonly used form of eosin.
    • The staining procedure for H&E follows a basic protocol including dewaxing, rehydration, haematoxylin staining, differentiation, and counterstaining with eosin, among other steps.
    • The balanced coloration of the stained specimen is the result of the intensity of the alum-hematoxylin and eosin, with alum-hematoxylin able to stain cytoplasm and eosin Y able to stain nuclear basic protein.
    • The staining schedule for H&E includes various steps such as immersing the slide in xylene, washing with alcohol, staining with Harris' haematoxylin, differentiating with acid alcohol, and counterstaining with eosin, among others.
    • The microvilli on the columnar epithelium are very crisp in an example of H&E staining.

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    Description

    Test your knowledge of the mechanism of Hematoxylin and Eosin (H&E) staining with this quiz focused on the direct method of staining, the attraction between tissue elements and dyes, and the concept of basophilic and acidophilic components.

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