SOP--Standard Operating Procedure
MASSON (bright green method) staining, which is modified by mallory three-color staining, uses two or three anionic dyes to mix together or sequentially to complete the dyeing, which is related to the molecular size of the anionic dye and the permeability of the tissue. According to the different penetrating properties of the tissue, different anion dyes with different molecular sizes are selected for staining, and different tissue components can be displayed. It is mainly used for the differential staining of collagen fibers and muscle fibers. [Operating procedures]
1. Place paraffin sections in an oven at 60 ° C for 1 to 2 hours.
2. Paraffin section conventional xylene, ethanol dewaxing to water,
3. Harlem hematoxylin staining for 10 minutes,
4. Return to blue under running water,
5. Acid Fuchsia-Li Chunhong-Orange Yellow G liquid staining for about 8min, the tissue can be observed to be red under the microscope,
6. Wash slightly with 0.2% acetic acid water.
7.5% phosphotungstic acid solution differentiation and mordant for 5 minutes or longer, observe the tissue differentiation to different degrees of red under the microscope,
Wash slightly with 8.0.2% acetic acid,
9. The bright green solution is stained for 15 minutes or longer. This step should be observed under a microscope, and the tissue can be stained with green.
Wash lightly with 10.0.2% acetic acid and lightly with 95% alcohol.
11. Dehydrated, transparent, sealed as usual
 
[Experimental results]:
Under the microscope, the nucleus was green lotus color; the glial fibers were bright green; the muscle fibers were orange-red.
 
[Problems and solutions in the experiment]:
1. Bright green dyeing time problem
Countermeasure: The staining time is related to the type of tissue and room temperature. The staining time in winter should be longer than that in summer. However, the content and distribution of collagen fibers in different specimens of different tissues or different tissues are different. The staining time.
 
[Customer's common questions and answers]
1. Why is the green color of collagen fibers in the tissues different?
Answer: This is related to the tissue type and the distribution of collagen fibers. The parts with more collagen fibers may be easier to stain. When the collagen fibers are just formed or less, the staining solution is more difficult to combine. It is also possible that different parts of the tissue have different affinity for the dye, resulting in large differences in color.