Weigert’s Iron Hematoxylin Solution
- Weigert’s Iron Hematoxylin is the powerful, acid-resistant, regressive nuclear stain made with Ferric Chloride + Hematoxylin. It is NOT for routine H&E. It is the mandatory nuclear stain for almost all Special Stains.
- Invented by Karl Weigert in 1904, it is the only hematoxylin that is resistant to acidic solutions, ferric chloride, and acid decalcifiers. This is why all other hematoxylins (Harris, Gill, Mayer’s) wash out in Special Stains, but Weigert’s does NOT.
- If the lab does Van Gieson, Verhoeff-Van Gieson (VVG), Masson’s Trichrome, Oil Red O, Mucicarmine, or Perl’s – the lab MUST use Weigert’s. No other hematoxylin will work.
- Why Harris, Gill & Mayer’s FAIL in Special Stains :
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1. All routine hematoxylins use Aluminum as mordant (Aluminum-Hematein lake). Aluminum lake dissolves in acidic pH.
2. Special Stains like Van Gieson contains Picric Acid (pH 1.5), Masson contains Ponceau-Acid Fuchsin in Acetic Acid (pH 2.0). So Aluminum hematoxylin gets decolorized completely – nuclei become colorless.
3. Weigert’s uses Iron as mordant (Ferric-Hematein lake). Iron lake is resistant to acid up to pH 1.0. So nucleus remains jet black even after 30 minutes in acidic counterstains.
4. Weigert’s is NOT optional – It is written in all CAP, AFIP & Culling’s Manual protocols as mandatory.
- 100% Acid-Resistant – Only Hematoxylin for Special Stains: Only Weigert’s survives acidic pH of Van Gieson (pH 1.5), Masson Trichrome (pH 2), Mucicarmine (pH 2.5). Harris, Gill II, Mayer’s completely decolorizes. •Weigert’s guarantees black nuclei even after 30 min in acid.
- True Jet Black Nuclei – Not Blue: Gives intense blackish-blue to black nuclei, not blue like Harris. Black nuclei gives best contrast with red collagen (Van Gieson), green/blue collagen (Masson), red mucin (Mucicarmine).
- Fresh Mix Technology – Always Active: Supplied as 2 stable solutions. Mix only when needed. Unlike Ehrlich’s or Harris which lose potency after 6 months, Weigert’s Solution A & B are stable for 18 months. Fresh mixing ensures zero metallic scum.
- Mercury-Free & US AFIP Compliant: No Mercuric Oxide. Formulation exactly as per AFIP Manual of Histotechnology, Bancroft, and Culling. Accepted by all NABL, CAP, NABH auditors for special stains.
- Mandatory for 6 Major Special Stains: Recommended protocol stain for – 1. Van Gieson for Collagen, 2. Verhoeff-Van Gieson (VVG) for Elastic Fibers, 3. Masson’s Trichrome, 4. Southgate Mucin / Mayer’s Mucicarmine, 5. Oil Red O & Sudan Black, 6. Perl’s Prussian Blue (Iron).
- Regressive Control = Crisp Detail: Because you over-stain then differentiate, you get perfect control. Ideal for small biopsies, decalcified bone sections (acid-decalcified tissue where Harris fails completely).
- Packing: Kit of 2 x 50ml / 2 x100ml /2×250 ml / 2×500 ml
- Iron Lake Technology for Acid Resistance :
- When you mix equal parts A + B just before use, you form Ferric-Hematein lake:
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Solution A = Hematoxylin in Alcohol
Solution B = Ferric Chloride + Conc. HCl in water
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Step 1 – Iron Lake Formation: Ferric Chloride oxidizes Hematoxylin to Hematein and simultaneously binds as a mordant. This forms a deep blue-black Ferric-Hematein lake. The HCl in Solution B ripens it instantly.
Step 2 – Intense Nuclear Binding: Ferric ions have highest affinity for nuclear chromatin phosphate groups. Stains nucleus intense blue-black in 5-10 minutes.
Step 3 – Acid-Proof Binding: Once bound, the Ferric-Chromatin bond is covalent-like and acid-resistant. Even if you put the slide in 5% Acid Alcohol or Picric Acid (Van Gieson), the black nucleus does NOT decolorize. This is the secret.
Step 4 – Regressive Stain: It over-stains intentionally, then you differentiate in Acid Alcohol (1% HCl in alcohol) for 5-10 seconds to get crisp black nuclei on clear background. This regressive method gives maximum nuclear detail.
- For Pathologists:
- For Lab Owners:
- Technical Specifications:
- Perfect Combination Kits:
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1. Diagnostic Black Nuclei: In Van Gieson, pathologist needs to see black nuclei vs red collagen vs yellow muscle. If nuclei fade (with Harris), collagen cannot be differentiated from muscle. Weigert’s black nuclei makes fibrosis grading, tumor stroma invasion reporting accurate.
2. Works on Decalcified & Acid-Treated Tissue: Bone marrow, bone tumor sections treated with Standard Decal /10% Formic Acid / Nitric Acid for decalcification lose aluminum-hematoxylin binding. Weigert’s Iron still stains perfectly. Only hematoxylin that works on decalcified bone.
3. NABL & CAP Mandatory Compliance: CAP checklist ANP.22950 states “For special stains involving acidic counterstains, an acid-resistant hematoxylin such as Weigert’s shall be used.” Using Harris for Van Gieson is audit failure.
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1. Zero Repeat Rate for Special Stains: Special stains are expensive, If nuclei fades with Harris, you repeat the whole stain. Weigert’s success rate 100% – saves money.
2. Long Shelf Life – No Wastage: Solution A & B separate stable 18 months at RT. Weigert’s never wastes.
3. One Hematoxylin for All Special Stains: Instead of trying Harris for everything and failing, one Weigert’s kit covers Van Gieson, Masson, VVG, Mucicarmine, Perl’s. Simplifies inventory.
| Specification | Details |
|---|---|
| Type | Regressive Iron Hematoxylin |
| Working Solution | Mix equal parts A+B fresh |
| Stable | 30 min |
| pH of Working Solution | ∼1.2 |
| Stain Type | Regressive, Acid-Resistant Ferric-Hematein Lake |
| Staining Time | 7-10 min |
| Differentiation | 1% Acid Alcohol |
| Color | Jet Black Nuclei |
| Shelf Life | 18 Months for Stock Solutions at RT |
| Grade | For Special Stains |
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1. Fibrosis Kit: Weigert’s Iron Hematoxylin + Van Gieson Stain
2. Liver / Kidney Kit: Weigert’s Iron Hematoxylin + Masson Trichrome Kit (3 Solutions)
3. Elastic Fiber Kit: Weigert’s Iron Hematoxylin + Verhoeff Stain + Van Gieson.
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