细菌是否有芽孢以及芽孢的形状和位置都是细菌的重要特征，细菌的芽孢壁比营养细胞的细胞壁结构复杂、致密、通透性低，着色和脱色都比营养细胞困难。芽孢有很强的抗热和抗化学物质的特性，当采用碱性染料并加热或延长染色时间时，使菌体和芽孢都能着色，再用蒸馏水冲洗脱去菌体颜色，但芽孢的颜色仍然可以保留，可以在显微镜下明显区分芽孢和营养体的形态。
阿拉丁芽孢染色液 (Moeller法) 主要由石炭酸染色液、Moeller脱色液、Moeller亚甲蓝染色液组成，芽孢会被染成红色，菌体呈蓝色，操作简单，结果判断更可靠，但应注意，观察芽孢应在成熟期，但也不能过久，否则只能看到芽孢，而营养体已经消失。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：
S1510446
| Component
| 3×20mL
| Storage
|
S1510446A
| 石炭酸染色液
| 20mL
| RT. Store in the dark.
|
S1510446B
| Moeller 脱色液
| 50mL
| RT
|
S1510446C
| Moeller亚甲蓝染色液
| 20mL
| RT |
自备材料：
1、酒精灯、载玻片、接种环、显微镜
2、蒸馏水
操作步骤(仅供参考)：
1、在洁净无油脂的载玻片上滴加1滴蒸馏水。
2、用接种环挑取少许巨大芽孢杆菌斜面的菌体，与蒸馏水混合。制成涂片，自然干燥。
3、在涂液处滴加石炭酸染色液，使细菌分布均匀，并用弱火加热，使染液冒蒸汽约5min。
4、冷后水洗，用Moeller脱色液脱色2min，用水轻轻冲洗。
5、用Moeller亚甲蓝染色液复染0.5min，用水冲洗。
6、镜检：干燥后镜检，观察芽孢的形状、大小，在菌体中的位置以及是否使菌体胀大。
染色结果：
附注：镜检应注意观察：
形态
| 圆形或卵圆形
|
位置
| 中央，末端，次末端
|
大小
| 菌体细胞是否膨大
|
注意事项：
1、玻片应洁净，无油污。
2、芽孢形成于生长发育期，观察芽孢应在成熟期，但也不能过久，否则只能看到芽孢，而营养体已经消失。
3、染色过程应小心操作，防止菌脱落。
4、固定时不宜用高热火焰固定。
5、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Whether a bacterium produces spores, along with the shape and location of those spores, are important identifying characteristics of the bacterium. The spore coat of bacteria is structurally more complex, compact, and less permeable than the cell wall of vegetative cells, making spore staining and decolorization more difficult to achieve than that of vegetative cells. Spores exhibit strong resistance to heat and chemical agents. When basic dyes are used in combination with heating or extended staining time, both the bacterial cells and spores can be stained. Subsequent rinsing with distilled water removes the stain from the vegetative cells, whereas the stain in the spores remains intact, allowing for clear morphological differentiation between spores and vegetative cells under a microscope.
Aladdin Spore Staining Solution (Moeller’s Method) consists primarily of carbol fuchsin staining solution, Moeller’s decolorizing solution, and Moeller’s methylene blue staining solution. With this reagent, spores are stained red and vegetative cells are stained blue. The method features simple operation and reliable result interpretation. However, it should be noted that spores should be observed during their mature stage, but not overly late in the growth cycle—otherwise, only spores can be visualized, as the vegetative cells would have already disintegrated. This reagent is intended for research use only and is not suitable for clinical diagnosis or any other purposes.
Product Components and Storage Conditions:
S1510446
| Component
| 3×20mL
| Storage
|
S1510446A
| Carbol Fuchsin Staining Solution
| 20mL
| RT. Store in the dark.
|
S1510446B
| Moeller’s Decolorizing Solution
| 50mL
| RT
|
S1510446C
| Moeller’s Methylene Blue Staining Solution
| 20mL
| RT |
Materials Required (Self-prepared):
1. Alcohol lamp, glass slides, inoculation loops, microscope
2. Distilled water
Operating Procedures (For Reference Only):
1. Place a drop of distilled water on a clean, grease-free glass slide.
2. Use an inoculation loop to pick a small amount of Bacillus megaterium from the slant culture, mix it thoroughly with the distilled water to prepare a smear, and allow it to air-dry.
3. Add carbol fuchsin staining solution to cover the smear evenly; heat gently over low flame until the staining solution produces steam, and maintain this state for approximately 5 minutes.
4. After cooling, rinse the slide with water. Decolorize with Moeller’s decolorizing solution for 2 minutes, then rinse the slide gently with water.
5. Counterstain with Moeller’s methylene blue staining solution for 0.5 minutes, followed by rinsing with water.
6. Microscopic examination: Air-dry the slide and observe under the microscope. Identify the shape, size and position of spores within the bacterial cells, as well as whether the spores cause swelling of the cells.
Staining Results:
Spores
| Red
|
Vegetative Cells
| Blue
|
Notes: The following points should be observed carefully during microscopic examination:
Morphology
| Round or oval
|
Position
| Central, terminal, subterminal
|
Size
| Whether the vegetative cells are swollen
|
Precautions:
1. The glass slides should be clean and free of grease.
2. Spores are formed during the growth and development stage. Observe the spores at their mature stage, but not too late—otherwise, only spores can be seen as the vegetative cells would have disintegrated.
3. Handle with care during the staining process to prevent the bacterial smear from detaching.
4. Avoid high-temperature flame fixation during the smear-fixing step.
5. For your safety and health, wear a lab coat and disposable gloves during the operation.
ZJ26F0433899
危险品化学品经营许可证（带存储）