The cell wall of Mycobacterium contains abundant lipids that surround the peptidoglycan layer, which generally renders Mycobacterium difficult to stain. Conventional staining methods require heating and extended staining time to facilitate dyeing of the bacteria. Once mycolic acid in Mycobacterium binds to the dye, it is difficult to decolorize with acid decolorizing solution, hence the name acid-fast staining. The most representative protocol is the Ziehl-Neelsen staining method for Mycobacterium tuberculosis, which is the recommended hot-staining method by the World Health Organization (WHO).
Aladdin Antacid Stain Solution (Auramine O-Rhodamine Fluorescence Method) is a type of fluorescent staining solution that does not require heating and is relatively safer than acid-fast hot-staining solutions. Its staining principle is as follows: after staining with Auramine O-Rhodamine 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, enabling faster detection of acid-fast bacilli. This reagent is for research use only and is not suitable for clinical diagnosis or other purposes.
Product Components and Storage Conditions:
A1510396
| Component
| 4×100mL
| Storage
|
A1510396A
| Auramine O-Rhodamine Staining Solution
| 100mL
| RT. Store in the dark.
|
A1510396B
| Acid Decolorizing Solution
| 2×100mL
| RT
|
A1510396C
| Counterstain Solution
| 100mL
| RT. Store in the dark. |
Self-prepared Materials:
Inoculation loop, glass slides, 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 a sufficient amount of Auramine O-Rhodamine Staining Solution to cover the smear, stain for 30 minutes in the dark, and rinse with water.
3. Decolorize with Acid Decolorizing Solution for 3 minutes until the smear no longer shows yellow color, then rinse with water.
4. Stain with Counterstain Solution for 2 minutes, rinse with water, blot the excess water gently, and air-dry naturally.
5. Observe the prepared slide under a fluorescence microscope.
Staining Results:
Acid-fast bacilli
| Bright yellow or orange-yellow
|
Non-acid-fast bacilli, cells, background
| Dark yellow
|
Precautions:
1. Tighten the reagent bottle caps immediately after each use to prevent reagent volatilization and contamination.
2. The fluorescence of Auramine O fades easily; please perform all operations as much as possible away from light.
3. All the above-mentioned reagents are irritating to the human body; please take appropriate protective measures.
4. For your safety and health, please wear a lab coat and disposable gloves during operation.
5. Please use the reagents as soon as possible after opening to avoid affecting the results of subsequent experiments.
ZJ26F0333473