Theoretical and Practical Aspects of Routine H&E Staining (Online CE Course)

(based on 161 customer ratings)

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

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.

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Continuing Education Credits

P.A.C.E.® Contact Hours (acceptable for AMT, ASCP, and state recertification): 1.5 hour(s)
Approved through 9/30/2026
Approved through 9/30/2026

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.

Customer Ratings

(based on 161 customer ratings)

Course Outline

Click on the links below to preview selected pages from this course.
  • Introduction
  • 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
      • Examples of Synthetic Dyes
      • True or False: The first natural dyes were made from aniline and are, therefore, sometimes referred to as aniline dyes.
      • Identify the following dyes as either synthetic or natural in origin.
      • 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?
      • True or False: The type of chromophore present on a dye molecule determines its color.
  • 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 their appropriate use.
      • True or False: One advantage of alcohol-based eosins over aqueous eosins is that they will stain slower and produce a broader range of shades.
      • Eosin stains the cytoplasm of cells as well as other tissue constituents. Which of the following would not be stained with eosin?
  • 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
      • Table 4: Staining Time Comparisons
      • General H&E Staining Procedures
      • When performing regressive H&E staining, slides need to be deparaffinized and _____________ before they will be able to take up the aqueous-based ...
  • Some Common Problems Encountered in H&E Staining
  • 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 medical 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 associate's 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 System Director of Anatomic Pathology for 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 29 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. It will discuss basic H&E staining mechanisms and the classification of biological stains. The material presented details the chemistry, diagnostic application, and problem-solving strategies for H&E stains. 

breast 40x_edit


Breast H&E S11-792a with arrows


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Fallopian Tube H&E with arrows


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liver bx 40x H&E with arrows


not blued_edit