细菌鞭毛是细菌的运动器官，幽门螺杆菌能够从强酸性的胃内腔穿过胃上皮细胞上的黏液层达到胃上皮细胞的中性环境，这就是鞭毛运动作用的很好例证，通过鞭毛染色可以观察到鞭毛形态、数量和鞭毛在菌体分布的位置，鞭毛数量和在菌体上的分布位置是鉴定细菌的重要依据之一。
阿拉丁鞭毛染色液 (Leifson法) 采用Leifson法，又称利夫森氏染色(Leifson’s flagella stain)法，试剂比较灵敏，操作简单，结果判断更可靠。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
产品组分及储存条件：
F1510198
| Component
| 3X50ml
| Storage
|
F1510198A
| Leifson Buffer
| 50 mL
| RT
|
F1510198B
| Leifson稀释液
| 50 mL
| RT
|
F1510198C
| 复红染色液
| 50 mL
| RT |
自备材料：
酒精灯、载玻片、蒸馏水、接种环、显微镜
操作步骤(仅供参考)：
1、在洁净无油脂的载玻片上滴加2滴蒸馏水。
2、用接种环挑取无菌蒸馏水，再与血平板上菌落接触，允许细菌游到接种环蒸馏水中，再将接种环移到玻片上蒸馏水顶部轻点2次。
3、轻轻摇动玻片，使细菌分布均匀；切勿研磨和搅动，以防鞭毛脱落。
4、置室温或35°C恒温箱内干燥固定。
5、滴加刚配制的Leifson染色液于载玻片上，染色3～10min，当染色液的一半以上面积出现金属光泽膜时，用蒸馏水轻轻冲洗染色液，自然干燥。
6、镜检：从涂片边缘开始，由外及里，逐渐移至中心；细菌分布少的地方，鞭毛容易观察；细菌密集的地方，鞭毛被菌体挡住，不易观察。
染色结果：
菌体和鞭毛呈不同程度的红色。
注意事项：
1、玻片应洁净，无油污。
2、染色的菌种应连续传代多次，处于生长活跃期。
3、染色过程应小心操作，防止鞭毛脱落。
4、配制好的染色液不宜久置，尽量在2h内使用。
5、固定时不宜用高热火焰固定。
6、为了您的安全和健康，请穿实验服并戴一次性手套操作。
7、试剂开封后请尽快使用，以防影响后续实验效果。
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.
ZJ26F0333149
危险品化学品经营许可证（带存储）