BacterStain™ Auramine Solution

A fluorescent dye used in microbiology for the detection of acid-fast bacteria (AFB) through fluorescence microscopy
Code: 04012
Packaging: 100ml/bottle or according to the customer’s request..
Storage Conditions: 18 – 25°C.
Transportation: Ambient temperature.
Shelf Life: 24 months from the date of manufacture.

INTENDED USE

BacterStrain™ Auramine Solution is primarily used in the Auramine-O staining technique to identify acid-fast bacteria like Mycobacterium tuberculosis in clinical samples, such as sputum. This staining method enhances the sensitivity of detection, making it easier to identify these bacteria under ultraviolet (UV) light.

PRINCIPLES

BacterStrain™ Auramine Solution binds to the mycolic acids in the cell wall of acid-fast bacteria, causing them to fluoresce bright yellow or orange under UV light. This fluorescence makes it possible to detect acid-fast bacteria quickly and efficiently, aiding in the diagnosis of tuberculosis and other mycobacterial infections..

For Sputum:
- Prepare the sample: Use a swab to transfer a portion of sputum containing mucus (including any yellowish material) onto a glass slide and spread it into a thin smear. The recommended area for the smear is about ½ x 1 inch (or 2 cm²).
- Create the smear: Spread the sample using a circular motion to ensure an even distribution. Ensure the smear is of the proper thickness. Smears that are too thick may peel off during staining and may be difficult to decolorize, while smears that are too thin may not contain enough sample.
- Allow the sample to air dry: Let the smear dry naturally in the air before proceeding.
For Urine:
- Prepare the sample: Create a smear from three centrifuged morning urine sediment samples. After spreading the sample, allow it to dry naturally.
- Heat fixation: Fix the smear by using an electric slide warmer, which is the recommended method. Alternatively, the smear can be heat-fixed by passing the dried slide (with the smear facing up) through the blue cone of a flame 2-3 times.
Staining:
- Stain with Rhodamine-Auramine: Place the fixed slide onto a staining rack and immerse it in rhodamine-auramine solution for 15 minutes. Make sure the surface does not dry out during the staining process.
- Remove excess color: After staining, rinse the slide with distilled water to remove excess dye.
- Fluorescence decolorization: For 2-3 minutes, immerse the slide in acid-alcohol solution (fluorescence decolorizer).
- Rinse thoroughly: Wash the slide with distilled water to remove any remaining acid-alcohol solution.
- Stain with methylene blue: Immerse the sample in methylene blue solution for 3-4 minutes, ensuring the slide does not dry out.
- Rinse and dry: Rinse with distilled water and allow the slide to dry naturally.
Examine under the microscope: Check the slide under a fluorescence microscope using an excitation filter (K530) and a barrier filter (BG 12), or using an excitation filter (G-365) and a barrier filter (LP 420). The slide can be examined at high magnification (400X) and confirmed by immersion oil.

RESULTS

Positive Test: AFB will fluoresce bright orange-red against a black background.
Negative Test: Non-acid-fast bacteria will not fluoresce or may appear pale yellow, contrasting completely with the bright and acid-fast organisms.

BacterLab quality control of each batch of products.

STT	Evaluation criteria	Result
1	Specimen size	Size 1x2cm, oval owl located in the middle of the glass slide
2	Smoothness	The specimen surface is continuous, even, not hollow or bare. Microscopic examination: Continuous microscopic fields without many empty microscopic fields, uniform brightness.
3	Thickness	Standard specimens have a thickness of about 0.04 mm. When the specimen is dry and not stained, it can be checked by placing a piece of paper with printed letters 4-5 cm below the specimen. If the letters are faint and readable, it is qualified. If the letters are illegible, it is thick. If the letters are too clear, it is thin.