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 around the gastric epithelial cells, which is a good example of the motile 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.
Aladdin Flagella Stain Solution adopts the Modified Ryu Staining Method. The advantage of this staining method is that it uses crystal violet as the core dye, which makes the reagent relatively stable, the operation simple, and the result interpretation more reliable. This reagent is intended for research purposes only and is not suitable for clinical diagnosis or any other applications.
Materials to Be Prepared:
Distilled water, inoculation loop, alcohol lamp, incubator, microscope, glass slide
Operating Procedures (For Reference Only):
1. Prepare the flagella staining working solution: Mix Reagent (A) and Reagent (B) at a ratio of 10:1 before use to obtain the flagella staining working solution. Store it at room temperature for later use.
2. Place 2 drops of distilled water onto a clean, grease-free glass slide.
3. 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.
4. Shake the glass slide slightly to distribute the bacteria evenly; do not grind or stir vigorously to prevent flagella detachment.
5. Air-dry and fix the smear at room temperature or in a 35°C incubator.
6. Add the flagella staining working solution dropwise and stain for 10–15 minutes.
7. Rinse the slide gently with water and air-dry naturally.
8. 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:
Both bacterial cells and flagella stain purple, with the color of bacterial cells being deeper than that of flagella.
Precautions:
1. Avoid flame fixation during the fixing step.
2. For your safety and health, wear a lab coat and disposable gloves during operation.
3. Use the reagent as soon as possible after opening to prevent it from affecting the results of subsequent experiments.
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