Theoretical and Practical Aspects of Routine H&E Staining (by LabCE)

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

(based on 124 customer ratings)

Author: Anita Buchiane, HT(ASCP) QIHC
Reviewer: Carla Shoffeitt, MSM, HT(ASCP)

Course provided by LabCE.

This course will explore the history and application of hematoxylin and eosin (H&E) stains used in the clinical histology laboratory. Basic H&E staining mechanisms and the classification of biological stains will be discussed. The material presented details the chemistry, diagnostic application, and problem solving strategies for H&E stains.

See more courses in: Histology

Included In These Course Packages

Continuing Education Credits

P.A.C.E.® Contact Hours (acceptable for AMT, ASCP, and state recertification): 1.5 hour(s)
Course number 578-011-18, approved through 5/31/2020
Course number 20-658952, approved through 9/1/2020


  • 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.

Customer Ratings

(based on 124 customer ratings)

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?
  • Categories of Biological Stains
      • Three Broad Categories of Biological Stains
  • Classification of Biological Stains
      • Classification of Biological Stains
      • Origin of Dyes: Natural
      • Origin of Dyes: Synthetic
      • Examples of Synthetic Dyes
      • Chemistry of Dyes
      • Mechanisms of Action
      • Progressive or Regressive Hematoxylin Staining
  • 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
  • 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
  • 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
  • References
      • References

Additional Information

Level of instruction: Intermediate
Intended audience: Clinical laboratory histotechnologists and 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: Anita Buchiane, HT(ASCP) QHIC, is the Lead Histology Technician at Brattleboro Memorial Hospital in Brattleboro, Vermont. Prior to this position, she worked as a Neuropathology/Special Procedures Technician at Baystate Medical Center in Springfield, Massachusetts.  Ms. Buchiane is a graduate of Hartford Hospital School of Allied Health in Connecticut, where she earned her Certificate in Histotechnology, and Greenfield Community College, Greenfield, Massachusetts, where she earned an associates degree in Liberal Arts with a math and science concentration. She is a member of the National Society for Histotechnology and the VT/NH Society for Histotechnology.  She was the recipient of the 2007 Lee G. Luna Foreign Travel Scholarship awarded by the National Society for Histotechnology and the 2010 Region 1 Scholarship awarded by the VT/NH  Society for Histotechnology.
Reviewer information: Carla J. Shoffeitt, MSM, HT(ASCP) is the Manager of the Pathology Department of Emory St. Joseph’s Hospital of Emory Healthcare in Atlanta, Georgia. She holds a Master of Science Degree in Healthcare Management as well as a Bachelor of Science degree in Biology from Troy University, Troy, Alabama. She is certified as a Histotechnician and has 25 years of experience in the field of Histology.
Course Description: This course will explore the history and application of hematoxylin and eosin (H&E) stains used in the clinical histology laboratory. Basic H&E staining mechanisms and the classification of biological stains will be discussed. The material presented details the chemistry, diagnostic application, and problem solving strategies for H&E stains. 

How to Subscribe
Histology CE (by LabCE)
Package of 35 online courses$65
ASCLS members save $10
Add to cart
Single online course$20
Add to cart
  • Order for instant access through any computer, any browser.
  • No shipping, faxing, or waiting for certificates.
  • Print or save your certificate of completion as soon as you've completed the course.
  • You'll have 90 days to complete your purchased courses.
Course provided by LabCE.
breast 40x_edit

Breast H&E S11-792a with arrows


Fallopian Tube H&E with arrows


liver bx 40x H&E with arrows

not blued_edit

ASCLS CE's design and platform are provided by MediaLab, Inc.