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Questions and Answers
What is the primary reason for removing water from the stained section?
What is the primary reason for removing water from the stained section?
What is the purpose of immersing the slide in a series of alcohol concentrations?
What is the purpose of immersing the slide in a series of alcohol concentrations?
Why is it important to observe the sections under the microscope after dehydration?
Why is it important to observe the sections under the microscope after dehydration?
What is the role of xylene in the clearing step?
What is the role of xylene in the clearing step?
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What is the purpose of raising the refractive index of the section?
What is the purpose of raising the refractive index of the section?
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What is the composition of the mounting agent DPX?
What is the composition of the mounting agent DPX?
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What should you do if you observe incomplete clearing or dehydration after mounting in DPX?
What should you do if you observe incomplete clearing or dehydration after mounting in DPX?
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What is the ultimate goal of the procedures after staining?
What is the ultimate goal of the procedures after staining?
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Why is Eosin Y the most widely used?
Why is Eosin Y the most widely used?
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What is the purpose of adding acetic acid to the eosin solution?
What is the purpose of adding acetic acid to the eosin solution?
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What is the first step in the H&E staining procedure?
What is the first step in the H&E staining procedure?
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Why is dewaxing necessary in the H&E staining procedure?
Why is dewaxing necessary in the H&E staining procedure?
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What is the purpose of hydration in the H&E staining procedure?
What is the purpose of hydration in the H&E staining procedure?
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What is the consequence of excessive heat during dewaxing?
What is the consequence of excessive heat during dewaxing?
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What is the role of alcohol in the H&E staining procedure?
What is the role of alcohol in the H&E staining procedure?
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What is the purpose of adding thymol to the eosin solution?
What is the purpose of adding thymol to the eosin solution?
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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.