Eosin Y in Staining
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Questions and Answers

What is the primary reason for removing water from the stained section?

  • Because water is not miscible with the mounting agent (correct)
  • To allow the mounting agent to bond with the tissue
  • To preserve the stained tissue
  • To increase the refractive index for viewing under the microscope
  • What is the purpose of immersing the slide in a series of alcohol concentrations?

  • To prevent distortion of the tissue (correct)
  • To preserve the stained tissue
  • To remove impurities from the tissue
  • To increase the refractive index
  • Why is it important to observe the sections under the microscope after dehydration?

  • To check for impurities in the tissue
  • To determine if additional differentiation is needed (correct)
  • To assess the level of dehydration
  • To check for mounting agent residue
  • What is the role of xylene in the clearing step?

    <p>To remove alcohol from the section</p> Signup and view all the answers

    What is the purpose of raising the refractive index of the section?

    <p>To allow for better visualization of tissue details</p> Signup and view all the answers

    What is the composition of the mounting agent DPX?

    <p>Dibutyl phthalate, Polystyrene, Xylene</p> Signup and view all the answers

    What should you do if you observe incomplete clearing or dehydration after mounting in DPX?

    <p>Return the slides to the clearing agent and re-mount</p> Signup and view all the answers

    What is the ultimate goal of the procedures after staining?

    <p>To preserve the stained tissue permanently</p> Signup and view all the answers

    Why is Eosin Y the most widely used?

    <p>Because it is soluble in alcohol</p> Signup and view all the answers

    What is the purpose of adding acetic acid to the eosin solution?

    <p>To sharpen the staining</p> Signup and view all the answers

    What is the first step in the H&E staining procedure?

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

    Why is dewaxing necessary in the H&E staining procedure?

    <p>To remove wax from the tissue section</p> Signup and view all the answers

    What is the purpose of hydration in the H&E staining procedure?

    <p>To prepare the tissue section for staining</p> Signup and view all the answers

    What is the consequence of excessive heat during dewaxing?

    <p>It damages the tissue</p> Signup and view all the answers

    What is the role of alcohol in the H&E staining procedure?

    <p>It removes the clearing agent from the tissue section</p> Signup and view all the answers

    What is the purpose of adding thymol to the eosin solution?

    <p>To inhibit the growth of fungi</p> Signup and view all the answers

    Study Notes

    Eosin Y

    • Eosin Y is the most widely used due to its solubility in alcohol.
    • It is typically used as a 0.5 or 1% solution in DI water with a crystal of thymol added to inhibit fungal growth.
    • Adding 0.5 ml of acetic acid to 1000 ml of stain solution can sharpen the staining.

    Staining Procedure for H&E

    • The procedure consists of 9 steps: dewaxing, hydration, hematoxylin, differentiation, bluing, eosin, dehydration, clearing, and cover-slipping.

    Preparation of Wax Sections for Staining

    • Dewaxing is necessary to remove wax from sections, as wax hinders the reaction of dyes with tissues and renders them impermeable to stains.
    • Xylene is used as a clearing agent, and heat can be incorporated to speed up the reaction.
    • Sections must be dry before immersing them in xylene.

    Hydration

    • Immerse sections in decreasing concentrations of alcohol (100% → 90% → 70% → water) to remove the clearing agent.
    • Replaced cloudy alcohol solutions with fresh ones.

    Procedures After Staining

    • It is essential to mount the stained section with a mounting agent to preserve the tissue and increase the refractive index for viewing under a microscope.

    Mounting Procedures

    • Dehydration: remove water from the stained section by immersing it in a series of alcohols (70% → 90% → absolute alcohol) to prevent distortion of the tissue.
    • Clearing: remove alcohol with xylene, which raises the refractive index of the section.
    • Mounting: use a mounting agent (DPX) to permanently preserve the stained section, ensuring no bubbles are trapped.

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    Description

    Learn about the properties and uses of Eosin Y, a popular stain in laboratory settings, including its solubility, concentration, and additives.

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