Thionin acetate (Thionine acetate) is a widely used metachromatic cationic dye in histological staining.

1. Cell staining
Experimental steps:
1. Prepare Thionin acetate solution: Dissolve Thionin acetate in an appropriate solvent (usually distilled water or PBS), at a concentration generally between 0.1% and 0.5%.
2. Fix the cells: Apply the cells to be stained onto the slide and fix the cells with an appropriate fixative (such as methanol or formaldehyde) after air-drying.
3. Staining: Add Thionin acetate solution dropwise to the sample. The staining time is usually 5-10 minutes and can be extended as needed.
4. Cleaning: Gently clean the sample with distilled water to remove excess dye.
5. Microscope observation: Use a microscope to observe the cell staining effect. Thionin acetate will produce a purple-blue staining effect to highlight the cell nucleus.
2. Staining tissue sections
Experimental steps:
1. Prepare Thionin acetate solution: Configure the solution as needed, the concentration is generally 0.1% to 0.5%.
2. Preparation of tissue sections: Fix the tissue sections on a slide, fix them with fixative (such as 10% formalin solution), and clear them after conventional dehydration.
3. Staining: Add Thionin acetate solution dropwise to the sections. The staining time is usually 5-15 minutes. The time can be adjusted appropriately depending on the tissue type.
4. Clean and dehydrate: Clean the sections with distilled water and then dehydrate.
5. Seal and observation: Seal the tablet with an appropriate sealant, and then observe the staining effect under a microscope.
3. Neural tissue staining:
Experimental steps:
1. Prepare Thionin acetate solution: The solution concentration is usually 0.1% to 0.5%.
2. Brain tissue section processing: Perform routine fixation, sectioning and dehydration of brain tissue.
3. Staining: Add Thionin acetate solution dropwise to the sections. The staining time is usually 10-20 minutes, which is suitable for observing the details of the neural tissue.
4. Clean and seal the sections: Wash the sections with water or PBS, then seal the sections and observe the brain tissue structure using a microscope.
Notes:
1. Concentration control: Too high concentration of Thionin acetate may lead to excessive staining, so the staining concentration needs to be adjusted according to experimental needs.
2. Staining time: The staining time should be optimized according to the type of tissue and experimental needs to avoid excessive staining or excessive background due to excessive staining time.
3. Tissue pretreatment: Before performing Thionin acetate staining, it is necessary to ensure that the tissue sections are properly fixed and dehydrated to ensure the staining effect.
4. Microscopic observation: Use an optical microscope to observe the sample. The nucleus stained by Thionin acetate is purple-blue, which is suitable for observing cell and tissue structure.