Course Outline
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- 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
- Types of eosin
- Purpose of Eosin
- Eosin Y
- Eosin-Phloxine
- EA Family (50 and 65)
- Differentiators
- Bluing
- Progressive versus regressive
- Protocol Selection
- Understained Slides
- Common causes
- The role of acidity in hematoxylin
- Water and eosin
- Lower cellularity of the sample can be misleading
- Overstained Slides
- Common causes
- 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
- Uneven Staining
- Common causes
- Inadequate Deparaffinization
- Suboptimal Section Quality
- Water and eosin
- 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
- Floaters
- "Burnt" edges
- Proper Specimen Collection and Handling
- Related to Processing
- Pigmentation
- Other Staining Considerations
- Automated Platform versus Hand Staining
- Why Move from Hand Staining to an Automated Platform?
- Charged slides versus adding adhesives to waterbaths
- Xylene substitutes
- Xylene Versus Xylene Substitutes
- Laboratory Safety
- Are ethanol or reagent alcohols my only dehydrant options?
- Conclusions
- Common H&E challenges
- A few favorite protocols
- References
Additional Information
Level of Instruction: Basic to intermediate
Intended Audience: Clinical laboratory histotechnologists, technicians, and other medical laboratory personnel who have an interest in this subject matter. This course is also appropriate for histology and clinical laboratory science students, pathology residents, and practicing pathologists.
Author information: Cynthia Sampias, JD, CT(ASCP)HTL is currently a Senior Field Support Specialist with Leica Biosystems. Her primary focus is to assist histology teams on all core histology products/consumables and troubleshooting. During her career, she worked as a Histotechnologist and Global Project Manager for Covance Central Laboratory Services and as a Senior Cytotechnologist/Histotechnologist at Diagnostic Cytology Laboratories, both located in Indianapolis, Indiana. She holds a Doctor of Jurisprudence degree from Indiana University School of Law with a specialty in health care law, and a Masters of Public Affairs degree in Health Services Administration from Indiana University Northwest. She is certified as both a cytotechnologist and histotechnologist.
Reviewer information: Donna M. Hedger, BS HT(ASCP), is currently a sales specialist at MediaLab. After graduating from Georgia Southern University with a Bachelor of Science in Biology, she spent 25 years working in the operations of the healthcare laboratory at Northside Gwinnett (formerly Gwinnett Medical Center). During her tenure there, she worked in various roles, including phlebotomy, lab assistant, pathology assistant, histology technician and more recently as the manager of the Pathology Laboratory for 18 years. She is a certified histology technician.
Course description: This course is intended to examine common challenges with H&E staining and offer guidance for troubleshooting.
