Bacterial flagella are the motile organs of bacteria. Helicobacter pylori can pass through the mucus layer on gastric epithelial cells from the strongly acidic gastric lumen to reach the neutral environment of gastric epithelial cells, which is a good example of the motility function of flagella. Flagella staining allows the observation of flagellar morphology, quantity, and their distribution on bacterial cells. The number and distribution of flagella on bacterial cells are among the important criteria for bacterial identification.
Common flagella staining methods include the Modified Ryu Method, Carbol Fuchsin Method, Leifson Method, Silver Staining Method, Löffler Method, and Cerares-Gill Method. Aladdin Flagella Stain Solution (Cerares-Gill Method), also known as Cerares-Gill Staining Method, is mainly suitable for bacteria with flagella that are difficult to stain, plant pathogenic bacteria that produce large amounts of gelatinous substances, and bacteria with very thin flagella that are prone to detachment. This reagent is intended for research purposes only and is not suitable for clinical diagnosis or other applications.
Product Components and Storage Conditions:
F1510195
| Component
| 2×50ml
| Storage
|
F1510195A
| Mordant solution A
| 25 mL
| RT
|
F1510195B
| Mordant solution B
| 15mL
| RT
|
F1510195C
| Mordant solution C
| 10 mL
| RT
|
F1510195D
| Fuchsin counterstain solution
| 50 mL
| RT. Store in the dark. |
Immediately before use, mix reagents A, B and C thoroughly at a volume ratio of 5:3:2 to prepare the mordant solution. Filter the solution properly prior to staining, and dilute it with 50%–100% water depending on the staining results. Do not prepare the solution in advance.
Materials to Be Prepared:
Alcohol lamp, glass slide, inoculation loop, microscope, distilled water
Operating Procedures (For Reference Only):
1. Prepare a smear of the bacterial suspension, then fix it by heating or air drying.
2. Add mordant solution dropwise onto the smear and treat for 3–5 minutes.
3. Rinse the smear with running water to remove the mordant solution.
4. Add fuchsin counterstain solution dropwise and treat for 3–5 minutes.
5. Rinse with water and air dry.
6. Microscopic examination: Start from the edge of the smear and move gradually from the periphery to the center. Flagella are easier to observe in areas with sparse bacterial distribution; in densely populated bacterial areas, flagella are obscured by the bacterial cells and thus difficult to observe.
Staining Results:
Bacterial cells and flagella show varying degrees of red color.
Precautions:
1. Glass slides should be clean and free of oil stains.
2. The strains to be stained should be subcultured continuously for multiple generations and in the logarithmic growth phase.
3. Handle with care during the staining process to prevent flagella detachment.
4. The mordant solution should not be stored for a long time; it is best prepared fresh before use.
5. Avoid high-temperature flame fixation during the fixing step.
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.
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