Masson staining kit

2-8 ℃, avoid light

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!

a year

microscope

1. Paraffin section
2. Frozen section

1. Microscope
2. 37 ℃ temperature chamber
3. Pipette
4. Refrigerator
5. Ice box

1. Fixed liquid: Choose formaldehyde or formaldehyde salt solution
2. Distilled water
3. Series Ethanol

1. Centrifugal tube
2. Suction head
3. Disposable gloves

1. Mix equal amounts of A1 and A2 to obtain Weigert's hematoxylin staining solution, which usually loses its staining power after 24 hours.
2. Each staining step should keep the slide moist.
3. Slice dewaxing should be as clean as possible.
4. Fixation plays an important role, and using different fixatives can prolong or shorten staining time.
5. The differentiation time of reagent B should vary depending on the thickness of the slice and the type of tissue, whether it is old or new.
6. The differentiation of reagent F phosphomolybdic acid should be controlled under the microscope until the collagen fibers appear light red or red. The differentiation time depends on the depth of staining, usually 1-2 minutes.
7. If the blue staining of collagen fibers is too light during microscopic observation, a counterstain solution can be used to counterstain for 2-3 minutes until the color is suitable. The dyeing time of the counterstain should not be too long, otherwise the color will be too dark and the contrast will not be strong.
If the overall dyeing effect is not good, soak in water for 60-90 minutes, fade, and then repeat the dyeing.

Reagent preparation:
Mix A1 and A2 evenly before use to obtain dye solution A. Use within 24 hours and do not prepare in advance.

Pre staining treatment of samples:
paraffin section
1. Dewaxing: dewaxing in xylene for 10-15 minutes. Replace with fresh xylene dewaxing for 10-15 minutes.
2. Soak the dewaxed film in 95% ethanol for 2 minutes, and then use 70% and 30% ethanol for 2 minutes each.
3. Hydration: Soak in distilled water for 2 minutes.
4. Rinse: Rinse twice with water at 30-40 ℃ for 30-60 seconds each time.

Staining of samples
1. Wet the glass slide with distilled water for 30-60 seconds.
2. Stain with freshly prepared staining solution A for 5-10 minutes.
3. Color separation liquid B separates colors for 6-8 minutes. Wash with water.
4. Treat with reagent C for 2-3 minutes.
5. Wash with distilled water for 1-2 minutes.
Stain with staining solution D for 5-10 minutes.
7. Prepare a weak acid working solution with distilled water: weak acid E=2:1, and wash with the weak acid working solution for 1 minute.
8. Wash reagent F for 1-2 minutes. It needs to be observed under a microscope and processed until the collagen fiber city is light red; Fiber red is sufficient.
9. Wash with weak acid working solution for 1 minute.
10. Stain with dye solution G aniline blue for 1-2 minutes.
11. Wash with weak acid working solution for 1 minute.
12. Rapid dehydration of 95% ethanol.
Dehydrate anhydrous ethanol three times, each time for 5 seconds.
14. Xylene is transparent three times, each time for 1 minute.
15. Seal the sheet with a sealing agent.
16. Observation of Results.

Result:
The nucleus, collagen fibers, mucus, and cartilage are blue in color;
The cytoplasm, muscle, red blood cells, cellulose, and nerve collagen are red in color.

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