太鼎生物科技/太鼎食安科技

Masson's three-color staining | 膠原纖維染色Servicebio貨號G1006-100mL(6×100 mL)

產品介紹:

Masson's三色染色法，又稱Masson's染色，是比較經典的膠原纖維染色法(collagen fiber staining method)。該方法利用結締組織(connective tissues)中陰離子生物染料(anionic biological dyes)和分子量(molecular weights)的不同滲透性。分子量小的染料很容易穿透結構緻密、滲透性低的組織，而分子量大的染料只能進入結構疏鬆、滲透性好的組織，因此不同的組織成分呈現出不同的顏色。

本品為試劑盒，主要成分如下：A液為2.5%重鉻酸鉀(2.5% Potassium dichromate)，B液、C液等體積混合為魏格鐵蘇木精染液(Weigert iron hematoxylin dye solution)，D液為麗春紅酸品紅(Ponceau acid fuchsin)，E液為1 %磷鉬酸溶液(phosphomolybdic acid solution)，F液為2.5%苯胺藍溶液(2.5% aniline blue solution)。結締組織切片(connective tissue sections stain)經Masson's三色染色液染色後，膠原纖維(collagen fibers)呈天藍色至亮深藍色，肌纖維(muscle fibers)、細胞質(cytoplasm)、纖維素(cellulose)、角質白(keratinous white)呈紅色至紫紅色，紅血球細胞(red blood cells)呈淡紅色。

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 Asolution 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.

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

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