7. Masson Tricolor Staining Kit; 100 mL×6 $400

Product Information

Description

Masson’s three-color staining method, also known as Masson staining, is a more classic collagen fiber staining method. This method makes use of the different permeability of anionic biological dyes and molecular weights in connective tissues. Dyes with small molecular weights can easily penetrate tissues with dense structures and low permeability, while dyes with large molecular weights can only enter tissues with loose structures and good permeability, so different tissue components present different colors.

This product is a kit, the main components are as follows: A solution is 2.5% Potassium dichromate, B solution and C solution are mixed in equal volume to be Weigert iron hematoxylin dye solution, D solution is Ponceau acid fuchsin, E Liquid is 1% phosphomolybdic acid solution, and F liquid is 2.5% aniline blue solution. After the connective tissue sections stain with Masson’s trichrome staining solution, collagen fibers show sky blue to bright dark blue, muscle fibers, cytoplasm, cellulose, and keratinous white show red to purplish red, and red blood cells show light red.

Storage and Handling Conditions

Store at room temperature, valid for 12 months.

Component

Assay Protocol

1. Section preparation: paraffin sections are deparaffinized to water; frozen sections stored at -20°C need to be allowed to stand and return to room temperature.

2. Soak the slices in Masson A solution at room temperature overnight (about 15 hours).

3. Soak the slices in Masson A solution and incubate in a 65° oven for 30 mins. Wash with tap water for 30 s until the yellow color on the tissue fades. At the same time, put Masson D liquid and Masson F liquid in a 65° oven to preheat.

4. Mix Masson B solution and Masson C solution in equal volume ( prepared for current use, not pre-prepared and preserved ), slice into the mixed solution and soak for 1 min, then rinse with running water.

5. Put the slices into 1% hydrochloric acid alcohol (concentrated hydrochloric acid: absolute ethanol = 1:100) for differentiation for about 1 min,until the nucleus was gray-black, and the background was almost colorless or light gray.

6. Rinse with tap water and drain the excess water on the slices. Dip the slices into Masson D solution for 6 minutes. At this time, the tissues will appear bright red. If the red is too light, the staining time can be prolonged.

7. Drain the slices slightly (not dry slices), and soak in Masson E solution for about 1 min. This step is for differentiation. It is enough to differentiate until the collagen fibers are light red and the fibers are red. The time of E solution can be adjusted according to the degree of staining, usually 1-2 min.

8. After the section is slightly drained of Masson E solution, without water washing, it is directly stained into Masson F solution for 2-30 s.

9. The slices are rinsed and differentiated in three consecutive tanks with 1% glacial acetic acid, each tank is about 8 seconds.

10. The slices are dehydrated in successive three cylinders of absolute ethanol for about 5 s, 10 s, and 30 s. Then two cylinders of n-butanol were dehydrated for 30 s and 2 minutes in sequence, and finally two cylinders of xylene were used to be transparent, 5 minutes each time, and the sheets were sealed with neutral gum.

Note

1. The temperature is low in winter. It is recommended to use Masson A solution, Masson D solution and Masson F solution after heating, and the corresponding dyeing time also needs to be adjusted.

2. After treating the Masson A, the washing time of slices should not be too long, which will lead to the overall weak and dark dyeing effect..

3. Masson E solution can be reused. If the dye solution changes color obviously, it needs to be replaced with a new one.

4. Please wear lab coat and disposable gloves during operation.

For Research Use Only!

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