Solarbio Masson’s Trichrome Stain Kit

Kit Components

Introduction

The Masson Trichrome Stain Kit is designed for the study of connective tissue, muscle, and collagen fibers. It aids in distinguishing collagen from smooth muscle, which can appear similar under a microscope. Nuclei are stained with Weigert’s iron hematoxylin, and cytoplasm and muscle are then stained with Beibrich scarlet-acid fuchsia. After treatment with phosphotungstic and phosphomolybdic acid, collagen is demonstrated by staining with aniline blue. Rinsing in acetic acid after staining renders the colors more delicate and transparent. This kit is suitable for use on formalin-fixed, paraffin-embedded, or frozen sections.

Protocols (For reference only)

1. Dewax to distilled water.
2. Stain with Weigert’s Iron Hematoxylin Solution for 5-10 minutes.
3. Differentiate with Acid Alcohol Differentiation Solution for 10-15 seconds.
4. Blue in Bluing Solution for 2-5 minutes. Rinse in deionized water.
5. Stain with Ponceau-Acid Fuchsia Solution for 5-10 minutes. Rinse in deionized water.
6. Differentiate in Phosphomolybdic Acid Solution for 1-2 minutes or until collagen is not red.
7. Without rinsing, add Aniline Blue Solution to the section and stain for 1-2 minutes.
8. Place section in Acetic Acid Working Solution (Mix 1 part Acetic Acid solution and 2 parts deionized water) for 1 minute.
9. Dehydrate quickly in 95% ethanol, absolute ethanol, and transparent in xylene.
10. Seal with resinene.

Result

• Nucleus: Black
• Cytoplasm, Muscle fibers: Red
• Collagen: Blue

Note

1. Slice dewaxing should be as clean as possible.
2. Equal amounts of A1 and A2 are mixed to form Weigert iron hematoxylin staining solution, generally losing its dyeing ability within 24 hours.
3. Tissue fixation plays an important role. Different fixatives can prolong or shorten the staining time.
4. In classic Masson trichromatic staining, Harris hematoxylin was used for nuclear staining, but Weigert hematoxylin is used in this staining solution to achieve brighter colors.
5. The differentiation time of acid ethanol should be determined based on slice thickness, tissue type, and age.
6. Weak acid solution can enhance color clarity and brightness.
7. The differentiation of phosphomolybdic acid should be controlled under a microscope until collagen fibers are light red.
8. Masson Bluing Solution can be replaced by Scott bluing solution or 0.1-1% lithium carbonate solution.