Understanding the Different Types of Haematoxylin in Histology: Which One Should You Use?

Not all haematoxylin stains are the same—learn the differences and which is best for your histology research.

Introduction

Haematoxylin is a cornerstone of histology staining, particularly in the widely used Hematoxylin and Eosin (H&E) stain. As a basic dye that stains nuclei shades of blue to purple, haematoxylin reveals vital cellular details that help researchers examine tissue architecture, identify cell types, and track disease-related changes.

But did you know there are multiple formulations of haematoxylin, each with unique properties?

In this blog, we explore the main types of haematoxylin used in research histology, what distinguishes them, and when you should choose one over another—especially if you're planning your tissue work with a histology provider like LabNexus in London.

Why Are There Different Types of Haematoxylin?

Pure haematoxylin extracted from the logwood tree is not a stain by itself—it must be oxidised to haematein and combined with a mordant (such as aluminium or iron salts) to become active.

Different oxidising agents and mordants create different formulations, each with slightly different:

• Staining intensity

• Colour tone

• Stability

• Tissue compatibility

• Background staining effects

These differences are important in both routine and advanced histological analysis, especially when high-quality, reproducible staining is required for research.

Main Types of Haematoxylin in Histology

1. Harris Haematoxylin

Overview: One of the most commonly used haematoxylins, Harris is widely used in H&E staining for both diagnostic and research histology.

• Colour tone: Strong, crisp blue to purple

• Mordant: Aluminium

• Oxidant: Sodium iodate

• Best for: General nuclear staining; excellent for high-throughput sectioning and routine H&E

Note: Original formulations contained mercury as a preservative, but modern versions are mercury-free and safer.

2. Mayer’s Haematoxylin

Overview: A milder, progressive stain used when more controlled nuclear staining is required.

• Colour tone: Softer blue/purple

• Mordant: Aluminium

• Oxidant: Sodium iodate

• Best for: Immunohistochemistry (IHC), cytology, and when background staining needs to be minimised

Key Feature: Because it's progressive, overstaining is less likely, making it suitable for delicate or lightly fixed tissues.

3. Gill’s Haematoxylin (Gill I, II, III)

Overview: A series of haematoxylins with varying strengths, used for both cytology and histology.

• Gill I: Light staining, for cytology

• Gill II: Moderate staining, general tissue use

• Gill III: Strong staining, ideal for histology

• Mordant: Aluminium

• Oxidant: Sodium iodate

• Best for: Consistent nuclear staining in H&E, especially when a slightly reddish tone is preferred

Gill’s haematoxylin is stable and can be used without differentiation steps in many workflows.

4. Delafield’s Haematoxylin

Overview: An older formulation, used mostly in academic or classic staining protocols.

• Colour tone: Rich violet-blue

• Mordant: Aluminium

• Oxidant: Natural (air/aging or chemical)

• Best for: Classic teaching or research protocols where deeper contrast is desired

Note: It requires aging to fully oxidise and is less commonly used in modern labs due to time and variability.

5. Weigert’s Haematoxylin

Overview: A special haematoxylin formulation using iron as a mordant instead of aluminium.

• Colour tone: Deep black-blue

• Mordant: Iron

• Oxidant: Ferric chloride

• Best for: Special stains like Masson’s Trichrome, Elastic Van Gieson (EVG), or stains where haematoxylin must resist acid solutions

Key Advantage: Weigert’s is more resistant to acidic counterstains, making it ideal for complex multicolour staining.

Choosing the Right Haematoxylin for Your Research

Depending on your tissue type, staining goals, and downstream applications, your choice of haematoxylin matters. Here's a simplified guide:

Research Use Case	Recommended Haematoxylin
General H&E staining	Harris or Gill III
Immunohistochemistry	Mayer’s
Trichrome and acid-resistant stains	Weigert’s
Cytology or light nuclear detail	Gill I or II
Teaching/classic protocols	Delafield’s

If you're using histology services in London, like those at LabNexus, our team can advise on the optimal stain based on your project and provide customised slide preparation.

LabNexus: High-Quality Haematoxylin Staining Services in London

At LabNexus, we specialise in research histology services, including:

• H&E staining with Gill, Harris, or Mayer’s haematoxylin

• Special stains requiring Weigert’s haematoxylin

• Full support for mouse and human tissues

• Custom slide quantities, orientations, and digital scanning

We work with researchers across London and the UK, offering fast turnaround, affordable pricing, and high-quality results you can trust.

Note: LabNexus only handles research samples—we do not offer diagnostic histology.

Book your free consultation to plan your next tissue staining project.

References

1. Bancroft, J.D., & Gamble, M. (2020). Theory and Practice of Histological Techniques (8th ed.). Elsevier.

2. Kiernan, J.A. (2008). Histological and Histochemical Methods: Theory and Practice (4th ed.). Scion Publishing.

3. Fischer, A.H., et al. (2008). Hematoxylin and eosin staining of tissue and cell sections. Cold Spring Harbor Protocols, 2008(5).

4. Suvarna, S.K., Layton, C., & Bancroft, J.D. (2018). Bancroft's Theory and Practice of Histological Techniques. Elsevier.