1 P.A.C.E. contact hour(s)

(based on 117 customer ratings)

See more courses in: Histology

Continuing Education Credits

P.A.C.E.® Contact Hours (acceptable for AMT, ASCP, and state recertification): 1 hour(s)
Course number 578-011-20, approved through 12/31/2021
Course number 20-767084, approved through 9/1/2022


  • Review the basics of H&E staining and what optimal staining should look like.
  • Identify common problems encountered with staining techniques.
  • Consider solutions to common artifacts that influence stain results.
  • Discuss the advantages of staining platforms and the use of kits to standardize stain performance.
  • Address outside factors that can influence H&E stain success.
  • Identify strategies used for successful H&E staining.

Customer Ratings

(based on 117 customer ratings)

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Course Outline

  • H&E Basics
    • Why is H&E the standard?
      • Introduction
      • Preferred by Pathologists for Routine Diagnosis
      • Ease of Use and Reproducibility
    • Making reagents versus purchasing ready to use
      • Making Staining Reagents in the Laboratory
      • Purchasing Ready to Use Reagents
      • Author's Thoughts
    • Types of hematoxylin
      • Purpose of Hematoxylin
      • Components of Hematoxylin
      • What are Mordants?
      • Mayer's Hematoxylin
      • Harris' Hematoxylin
      • Gill's Hematoxylin
      • Iron Hematoxylins
    • Types of eosin
      • Purpose of Eosin
      • Eosin Y
      • Eosin-Phloxine
      • EA Family (50 and 65)
    • Differentiators
      • Purpose of Differentiating Solutions
    • Bluing
      • Bluing Solutions
    • Progressive versus regressive
      • Progressive and Modified Progressive Staining Methods
      • Regressive Staining Method
  • Protocol Selection
    • Selecting a protocol
      • Does One Staining Protocol Fit All?
      • A Good Baseline Staining Protocol
      • How Best to Optimize Staining Protocol
    • Automated Platform versus Hand Staining
      • Why Move from Hand Staining to an Automated Platform?
  • Understained Slides
    • Common causes
      • Poor Coloration: Is the Protocol Long Enough?
      • One Stain Component Overshadowing Another
      • Is the Differentiating Solution the Right Intensity?
    • The role of acidity in hematoxylin
      • The Importance of pH
    • Water and eosin
      • Eosin Differentiation
    • Lower cellularity of the sample can be misleading
      • Where is the Pink and Blue?
  • Overstained Slides
    • Common causes
      • Protocol is Too Long
      • One Stain Component Under Staining Compared to the Other
      • Is the Differentiator Too Strong or the Step Too Long?
    • The role of acidity in differentiation of hematoxylin
      • Acid Extends Hematoxylin Life: Why do I Need it for Differentiation?
    • Water and eosin
      • The Impact of Water on Eosin
    • Tissue considerations
      • Cellularity Makes a Difference
  • Uneven Staining
    • Common causes
      • Inadequate Deparaffinization
      • Suboptimal Section Quality
    • Water and eosin
      • Impact of Excess Water Following Eosin
      • Water in Xylene
    • Water quality
      • Tap Water versus Deionized Water
  • Staining Frozen Tissues
      • Challenges with Frozen Tissue Staining
  • Artifacts That Can Make Staining a Challenge
    • The impact of suboptimal tissue processing
      • Sample Collection
      • Sample Fixation
      • Choosing Processing Protocols that are Appropriate for your Tissues
    • Nuclear bubbling
      • Nuclear Bubbling: Created During Processing
      • Nuclear Bubbling: Created After Cutting
    • Floaters
      • Where are the Floaters with Respect to the Tissue?
      • Fungal and Bacterial Contamination
    • "Burnt" edges
      • Proper Specimen Collection and Handling
      • Related to Processing
    • Pigmentation
      • Causes of Pigmentation
  • Other Staining Considerations
    • Charged slides versus adding adhesives to waterbaths
      • Adhesives
      • Charged Slides
      • Charged Slides Used with Adhesives
    • Xylene substitutes
      • Xylene Versus Xylene Substitutes
      • Laboratory Safety
    • Are ethanol or reagent alcohols my only dehydrant options?
      • Methanol, Isopropyl Alcohol, and Flex
  • Conclusions
    • Common H&E challenges
      • Change Early, Change Often!
      • Maintenance and Good Quality Control
      • Ensure Training
    • A few favorite protocols
      • Well-Rounded, Standard H&E - Regressive Protocol
      • Well-Rounded, Standard H&E - Modified Progressive Protocol
      • Well-Rounded, Standard H&E - Progressive Protocol
      • Adjusting for Coloration
  • References
      • References
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