Course Outline
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- H&E Basics
- Why is H&E the standard?
- Making reagents versus purchasing ready to use
- Types of hematoxylin
- Types of eosin
- Differentiators
- Purpose of Differentiating Solutions
- Bluing
- Progressive versus regressive
- Progressive and Modified Progressive Staining Methods
- Regressive Staining Method
- Protocol Selection
- Understained Slides
- Common causes
- The role of acidity in hematoxylin
- Water and eosin
- 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
- Water and eosin
- The Impact of Water on Eosin
- Tissue considerations
- Cellularity Makes a Difference
- Uneven Staining
- Common causes
- 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
- Nuclear bubbling
- Floaters
- Where are the Floaters with Respect to the Tissue?
- Fungal and Bacterial Contamination
- "Burnt" edges
- Pigmentation
- Other Staining Considerations
- Automated Platform versus Hand Staining
- Charged slides versus adding adhesives to waterbaths
- Adhesives
- Charged Slides
- Charged Slides Used with Adhesives
- Xylene substitutes
- 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
- 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.
