分枝杆菌的细胞壁内含有大量脂质包围在肽聚糖的外面，所以分枝杆菌一般不易着色。传统的染色方法要经过加热和延长染色时间来促使其着色。分枝杆菌中的分枝菌酸与染料一旦结合后，就很难被酸性脱色液脱色，故名抗酸染色。其中最具代表性是结核杆菌 Ziehl－Neelsen 染色法，该法是WHO推荐热染的方法。抗酸染色液 (金胺O荧光法) 属于荧光染色液，无需加热，相对较抗酸热染液安全。其染色原理是在室温条件下Auramine O 染色以及复染后，用含有紫外光源的荧光显微镜检查，抗酸杆菌呈亮黄色，而其他细菌及背景中的物质呈暗黄色，这种方法可用低倍镜检，因此能更快速找出抗酸性菌。
产品组分及储存条件：
A1510389
| Component
| 4×100mL
| Storage
|
A1510389A
| 金胺 O 染色液
| 100mL
| RT. Store in the dark.
|
A1510389B
| 酸性脱色液
| 2×100mL
| RT
|
A1510389C
| 复染液
| 100mL
| RT. Store in the dark. |
试验所需自备的仪器及材料：
接种环、载玻片、蒸馏水、荧光显微镜
操作步骤 (仅供参考)：
1、 接种环挑取待检样本，涂布于载玻片上，加热固定。
2、 滴加金胺 O 染色液，避光染色 10～15min，水洗。
3、 用酸性脱色液脱色 3～5min，直至涂片无黄色为止，水洗。
4、 用复染液染色 2min，水洗。
5、 轻轻吸干水分，自然干燥。
6、 荧光显微镜下镜检。
染色结果：
抗酸菌：亮黄色或橘黄色
非抗酸菌、细胞、背景：暗黄色
注意事项：
1、每次使用后盖紧试剂瓶，以防试剂挥发和污染。
2、金胺O光易衰减，尽量避光操作。
3、上述试剂均对人体有刺激性，请注意适当防护。
4、为了您的安全和健康，请穿实验服并戴一次性手套操作。
5、本产品仅由于科研，严禁他用。
The cell wall of mycobacteria contains a large amount of lipids surrounding the peptidoglycan layer, so mycobacteria are generally difficult to stain. Traditional staining methods require heating and prolonged staining time to facilitate dyeing. Once the mycolic acid in mycobacteria combines with the dye, it is difficult to be decolorized by acid decolorizing solution, hence the name acid-fast staining. The most representative one is the Ziehl-Neelsen staining method for Mycobacterium tuberculosis, which is the heat staining method recommended by the WHO. Acid-fast staining solution (Auramine O fluorescent method) belongs to the category of fluorescent staining solutions. It does not require heating and is relatively safer than acid-fast heat staining solution. Its staining principle is as follows: after Auramine O staining and counterstaining at room temperature, the specimen is examined under a fluorescence microscope equipped with an ultraviolet light source. Acid-fast bacilli appear bright yellow, while other bacteria and substances in the background appear dark yellow. This method allows examination under low magnification, thus enabling faster identification of acid-fast bacteria.
Product Components and Storage Conditions:
A1510389
| Component
| 4×100mL
| Storage
|
A1510389A
| Auramine O Staining Solution
| 100mL
| RT. Store in the dark.
|
A1510389B
| Acid Decolorizing Solution
| 2×100mL
| RT
|
A1510389C
| Counterstain Solution
| 100mL
| RT. Store in the dark. |
Instruments and Materials to Be Prepared by the User for the Test:
Inoculation loop, glass slide, distilled water, fluorescence microscope
Operating Procedures (For Reference Only):
1. Pick the sample to be tested with an inoculation loop, spread it evenly on a glass slide, and fix it by heating.
2. Add Auramine O staining solution dropwise, stain for 10–15 minutes away from light, and then rinse with water.
3. Decolorize with acid decolorizing solution for 3–5 minutes until the smear turns colorless, and then rinse with water.
4. Counterstain with counterstain solution for 2 minutes, and then rinse with water.
5. Gently blot away excess water and allow the smear to air-dry naturally.
6. Examine the smear under a fluorescence microscope.
Staining Results:
Acid-fast bacteria: Bright yellow or orange-yellow
Non-acid-fast bacteria, cells, and background: Dark yellow
Precautions:
1. Tighten the reagent bottle cap immediately after each use to prevent reagent volatilization and contamination.
2. Auramine O is prone to photobleaching; please perform all operations as far away from light as possible.
3. All the above-mentioned reagents are irritating to the human body; please take appropriate protective measures.
4. For your safety and health, wear a lab coat and disposable gloves during operation.
5. This product is intended for research use only and must not be used for any other purposes.
ZJ26F0333437
危险品化学品经营许可证（带存储）