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Shanghai Beibo Biotechnology Co., Ltd
Improved Masson staining kit
NegotiableUpdate on 05/06
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Overview
Product Overview: Connective tissue, in a narrow sense, refers to the three types of fibers it contains: collagen fibers, reticular fibers, and elastic fibers, with collagen fibers being the most distributed and abundant type of fiber. Masson trichrome staining, also known as Masson staining, is a method of staining connective tissue and a classic technique for staining collagen fibers. The so-called trichrome staining usually refers to staining the nucleus and selectively displaying collagen fibers and muscle fibers. The dyeing principle of this method is related to the size of anionic dye molecules and the permeability of tissues: the size of molecules is reflected by their molecular weight, and small molecular weights are easy to penetrate tissues with dense structures and low permeability; However, high molecular weight can only enter tissues with loose structure and high permeability. However, light green or aniline blue have high molecular weights, so after Masson staining, muscle fibers appear red, while collagen fibers appear green (light green) or blue (aniline blue), mainly used to distinguish between collagen fibers and muscle fibers. Bebo Improved Masson trichrome staining compared to conventional Masson trichrome staining .....
Product Details
storage temperature
2-8 ℃, avoid light
Precautions
1. The validity period is the period during which the reagent kit is stored under the required conditions before being opened.
2. Please use up the reagent as soon as possible after unpacking!
2. Please use up the reagent as soon as possible after unpacking!
Validity period
a year
detection method
microscope
Instrument preparation
1. 37 ℃ temperature chamber
2. Pipette
3. Refrigerator
4. Ice box
2. Pipette
3. Refrigerator
4. Ice box
Reagent preparation
1. 10% formalin fixative
2. Distilled water
3. Series Ethanol
2. Distilled water
3. Series Ethanol
Preparation of consumables
1. Centrifugal tube
2. Suction head
3. Disposable gloves
2. Suction head
3. Disposable gloves
Precautions for use
1. Slice dewaxing should be as clean as possible.
The differentiation time of acidic ethanol should be determined based on the thickness of the slices, the type and age of the tissue.
On the one hand, the function of phosphomolybdic acid is to differentiate collagen fibers stained with red into colorless or light red, while muscle fibers such as cellulose still appear bright red; On the other hand, it also has a mordant effect on collagen fibers, making them easier to bind with aniline blue solution.
4. After staining with aniline blue solution, treat with weak acid solution to remove the blue color from the original slurry, making the staining bright and clear. For tissues fixed with Zenker solution, weak acid solution treatment can be extended to 5 minutes.
5. Weak acid solution can make the color clearer and brighter. If used in large quantities, a 0.1-0.3% acetic acid solution can be prepared by oneself as a substitute.
The differentiation time of acidic ethanol should be determined based on the thickness of the slices, the type and age of the tissue.
On the one hand, the function of phosphomolybdic acid is to differentiate collagen fibers stained with red into colorless or light red, while muscle fibers such as cellulose still appear bright red; On the other hand, it also has a mordant effect on collagen fibers, making them easier to bind with aniline blue solution.
4. After staining with aniline blue solution, treat with weak acid solution to remove the blue color from the original slurry, making the staining bright and clear. For tissues fixed with Zenker solution, weak acid solution treatment can be extended to 5 minutes.
5. Weak acid solution can make the color clearer and brighter. If used in large quantities, a 0.1-0.3% acetic acid solution can be prepared by oneself as a substitute.
Usage
Operation steps (for reference only)
1. The tissue is fixed in 10% formalin fixative and routinely dehydrated and embedded. Slice thickness of 4um, conventional dewaxing to water;
2. Slice into reagent A and incubate at 37 ℃ for 2 hours for mordanting or overnight at room temperature.
3. Rinse with running water until the yellow color disappears.
4. Stain with reagent B for 2-3 minutes and wash with water.
5. Stain with reagent C for 2-3 minutes and wash with water.
6. Differentiate reagent D for a few seconds and rinse with running water for 10 minutes.
Stain with reagent E for 10 minutes and rinse with distilled water.
8. Treat with reagent F phosphomolybdic acid solution for 10 minutes.
9. Pour out reagent F without washing with water, and directly add reagent G for 5 minutes of aniline blue staining.
10. Treat with weak acid reagent H for 2 minutes.
11. 95% ethanol is rapidly dehydrated, and anhydrous ethanol is dehydrated three times, each time for 5-10 seconds.
12. Xylene should be transparent three times, each time for 1-2 minutes.
13. Neutral gum seal.
14. Observation of Results.
Result:
Collagen fibers: blue;
Muscle fibers, cytoplasm, cellulose, red blood cells, keratin: red;
Cell nucleus: Blue brown.
1. The tissue is fixed in 10% formalin fixative and routinely dehydrated and embedded. Slice thickness of 4um, conventional dewaxing to water;
2. Slice into reagent A and incubate at 37 ℃ for 2 hours for mordanting or overnight at room temperature.
3. Rinse with running water until the yellow color disappears.
4. Stain with reagent B for 2-3 minutes and wash with water.
5. Stain with reagent C for 2-3 minutes and wash with water.
6. Differentiate reagent D for a few seconds and rinse with running water for 10 minutes.
Stain with reagent E for 10 minutes and rinse with distilled water.
8. Treat with reagent F phosphomolybdic acid solution for 10 minutes.
9. Pour out reagent F without washing with water, and directly add reagent G for 5 minutes of aniline blue staining.
10. Treat with weak acid reagent H for 2 minutes.
11. 95% ethanol is rapidly dehydrated, and anhydrous ethanol is dehydrated three times, each time for 5-10 seconds.
12. Xylene should be transparent three times, each time for 1-2 minutes.
13. Neutral gum seal.
14. Observation of Results.
Result:
Collagen fibers: blue;
Muscle fibers, cytoplasm, cellulose, red blood cells, keratin: red;
Cell nucleus: Blue brown.
References
1. Yuyan Lu, et al.
Valsartan attenuates pulmonary hypertension via suppression of mitogen activated protein kinase signaling and matrix metalloproteinase expression in rodents
Molecular Medicine Reports 2017
2. Fei Hou, et al.
Attenuation of liver fibrosis by herbal compound 861 via upregulation of BMP-7/Smad signaling in the bile duct ligation model rat
Molecular Medicine Reports 2016
3. Huikuan Gao, et al.
Sequential changes in autophagy in diabetic cardiac fibrosis
Molecular Medicine Reports 2015
Valsartan attenuates pulmonary hypertension via suppression of mitogen activated protein kinase signaling and matrix metalloproteinase expression in rodents
Molecular Medicine Reports 2017
2. Fei Hou, et al.
Attenuation of liver fibrosis by herbal compound 861 via upregulation of BMP-7/Smad signaling in the bile duct ligation model rat
Molecular Medicine Reports 2016
3. Huikuan Gao, et al.
Sequential changes in autophagy in diabetic cardiac fibrosis
Molecular Medicine Reports 2015
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