There are various methods for staining mucous substances, such as AB-PAS staining, mucin HID-AB staining, standard Alcian blue staining, etc. Most of these methods utilize the principle that Alcian blue, a cationic dye, can bind to acidic groups-specifically, Alcian blue forms insoluble complexes with anionic groups (e.g., carboxyl and sulfate groups) contained in tissues. However, the aforementioned methods are not designed specifically for demonstrating neutral mucous substances.
Mucicarmine staining is mainly used to display neutral mucous substances; it can also stain acidic mucous substances and Cryptococcus neoformans. It is particularly suitable for showing mucus in the stomach and epithelium, and is clinically widely used for the differentiation of mucus-secreting epithelial carcinomas. This reagent is for research use only and not intended for clinical diagnosis or other purposes.
M1508531
| Component | 3×50 mL | Storage |
| M1508531A | LM Hematoxylin Staining Solution
| 50 mL | RT. Store in the dark. |
| M1508531B | Acidic Ethanol Differentiating Solution
| 50 mL | RT. |
| M1508531C | Mucicarmine Stock Solution
| 10 mL | 2-8℃. Store in the dark. |
Materials to Be Prepared by the User
1. Distilled water, graded ethanol/xylene or dewaxing-clearing solution, neutral gum
2. Microscope, glass slides
Operating Procedures (For Reference Only)
1. Dewax sections to water using xylene or dewaxing-clearing solution; cell smears should be fixed before washing with water.
2. Stain cell nuclei with LM Hematoxylin Stain Solution for 3-5 minutes (for cell smears, 1-3 minutes is sufficient), followed by rinsing with running water for 5 minutes.
3. Differentiate with Acidic Ethanol Differentiating Solution for 2-5 seconds, then rinse with running water for bluing.
4. Prepare the mucicarmine working solution in advance during the above steps at a ratio of mucicarmine stock solution: distilled water=1:4. Counterstain the samples with the mucicarmine working solution for 30 minutes to several hours; adjust the duration according to room temperature (for cell smears, 20-30 minutes is adequate).
5. Rinse with running water.
6. Dehydrate with graded ethanol, clear with xylene or dewaxing-clearing solution, and mount with neutral gum.
Staining Results
Mucous substances: Red
Cell nuclei: Blue
Precautions
1. Use 10% neutral formalin as the fixative.
2. Stain cell nuclei lightly with LM Hematoxylin Stain Solution.
3. The mucicarmine working solution is volatile and prone to precipitation; use a vertical staining jar for staining. If precipitation occurs, filter the solution before use.
4. For counterstaining after mucicarmine working solution staining, rapid staining with low-concentration light green for a few seconds is recommended.
5. The ethanol dehydration process also has a differentiating effect; strictly control the dehydration time.
6. For your safety and health, wear a lab coat and disposable gloves during operation.
7. Use the reagent as soon as possible after opening to avoid affecting subsequent experimental results.
