Bacterial flagella are the motile organs of bacteria. Helicobacter pylori can penetrate the mucus layer on gastric epithelial cells from the strongly acidic gastric lumen to reach the neutral microenvironment surrounding the gastric epithelial cells, which is a good example of the motile function of flagella. Flagella staining enables 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.
Aladdin Flagella Stain Solution (Leifson Method), which adopts the Leifson Method and is also known as Leifson’s Flagella Stain Method, features high sensitivity, simple operation, and more reliable result interpretation. This reagent is intended for research purposes only and is not suitable for clinical diagnosis or any other applications.
Product Components and Storage Conditions:
F1510198
| Component
| 3×50ml
| Storage
|
F1510198A
| Leifson Buffer
| 50 mL
| RT
|
F1510198B
| Leifson Diluent
| 50 mL
| RT
|
F1510198C
| Fuchsin Stain Solution
| 50 mL
| RT |
Materials to Be Prepared:
Alcohol lamp, glass slide, distilled water, inoculation loop, microscope
Operating Procedures (For Reference Only):
1. Place 2 drops of distilled water onto a clean, grease-free glass slide.
2. Use an inoculation loop to take a small amount of sterile distilled water, then touch the colonies on the blood agar plate to allow the bacteria to swim into the distilled water on the loop. Next, gently touch the top of the distilled water drops on the glass slide twice with the inoculation loop.
3. Shake the glass slide slightly to distribute the bacteria evenly; do not grind or stir vigorously to prevent flagella detachment.
4. Air-dry and fix the smear at room temperature or in a 35°C incubator.
5. Add freshly prepared Leifson stain solution onto the glass slide and stain for 3–10 minutes. When a metallic luster film appears over more than half of the stain solution area, rinse the stain solution gently with distilled water and air-dry naturally.
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 maintained in the logarithmic growth phase.
3. Handle with care during the staining process to prevent flagella detachment.
4. The prepared stain solution should not be stored for an extended period; use it as soon as possible within 2 hours.
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 the results of subsequent experiments.
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