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
| 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. |
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