Theoretical and Practical Aspects of Routine H&E Staining

Continuing Education Credits

• P.A.C.E.® Contact Hours: 1.5 hour(s)
• Florida Board of Clinical Laboratory Science CE Credit Hours -
  General (Histology): 1.5 hour(s)

Objectives

• Differentiate between natural and synthetic dyes.
• Analyze progressive staining versus regressive staining.
• Distinguish between the oxidants and mordants in common hematoxylin and eosin (H&E) stains.
• Identify cell constituents demonstrated with the H&E stain.
• Describe the purpose of and reagents used for deparaffinization, hydration, and dehydration.
• Identify potential problems encountered with routine staining and solutions to resolve them.

Course Outline

• Introduction
• Clinical Significance and Correlation of Histology
• How is Histology Useful to the Pathologist?
• Application of the Hematoxylin and Eosin (H&E) Stain
• Why is the H&E Stain the Most Common?
• Which of the following statements about histology are TRUE? (Choose all that apply.)
• Categories of Biological Stains
• Three Broad Categories of Biological Stains
• Which one of the following categories of stains is used to differentiate between a cell's nucleus and cytoplasm?
• Which category of stain would you employ if asked to demonstrate muscle in a tissue sample?
• Classification of Biological Stains
• Classification of Biological Stains
• An acidic dye has a pH less than 7 and has an affinity for __________ tissue components.
• Origin of Dyes: Natural
• Origin of Dyes: Synthetic
• The first natural dyes were made from aniline and are therefore sometimes referred to as aniline dyes.
• Examples of Synthetic Dyes
• Chemistry of Dyes
• Mechanisms of Action
• What is the staining mechanism that overstains tissue and then employs a differentiation step?
• Progressive or Regressive Hematoxylin Staining
• Which type of staining employs the process of differentiation to delineate desired tissue elements?
• Routine Staining in the Histology Laboratory
• Hematoxylin Oxidation
• Mordants
• Commonly used Hematoxylins: Their Oxidizers and Mordants
• Differentiating
• The Bluing Step
• Eosin as a Counterstain
• Types of Eosin
• Eosin Differentiation
• Match the following solutions with its' appropriate use.
• One advantage of using alcohol-based eosins over aqueous eosins is that they will stain slower and produce a wider range of shades.
• Other than the cytoplasm of cells, what other tissue constituents are stained with eosin? (Choose all that apply.)
• Applications of the H&E Stain
• Healthy Versus Diseased Tissue with H&E Stain
• Uterus
• Liver
• Breast
• Skin
• Appendix
• Fallopian Tube
• Additional Steps Employed in Routine H&E Staining
• Paraffin Sections
• Paraffin Sections, continued:
• Staining Time Comparisons
• General H&E Staining Procedures
• When performing regressive H&E staining, the slides must be deparaffinized and ___________ before they will be able to take up the aqueous-based h...
• Some Common Problems Encountered in H&E Staining
• Nuclear Staining Errors
• Poor Eosin Differentiation
• Poor Contrast Between the Hematoxylin and the Eosin
• Reddish- Brown Nuclei
• Dark Precipitate On The Slide
• Uneven Staining
• White Patches on Slides After Deparaffinization Step
• Clear Patches on Tissue After Hydrating
• Contaminated Clearing Agent
• Which of the following observations would indicate that tissue sections have not been sufficiently deparaffinized? (Choose all that apply.)
• What is the function of the hydration alcohols? (Choose all that apply.)
• What is the indication that the tissue did not go through the bluing step?
• References
• References

Additional Information