BacterStain™ Ziehl Neelsen Stain Kit

A set of reagents used for the dectection of acid-alcohol resistant bacteria.
Code: 04002
Packaging: Alcohol acid; Carbol fuchsin; Methylene blue: 3 bottles/box 100ml or as per customer request.
Storage Conditions: 18–25°C.
Transportation: Ambient temperature.
Shelf Life: 24 months from the date of manufacture.

INTENDED USE

BacterStrain™ Ziehl Neelsen Stain Kit can be used for the demonstration of Acid Alcohol fast bacilli.

PRINCIPLES

BacterStain™ Ziehl-Neelsen Stain Kit is based on the structural differences in the cell walls of certain Mycobacteria, which contain a high amount of mycolic acid. This makes it difficult for these bacteria to stain using traditional Gram staining methods. Therefore, the acid-fast staining method must be used to observe certain bacteria.
The Ziehl-Neelsen staining method uses two staining chemicals: carbol fuchsin and methylene blue, combined with a decolorizing agent (acid-alcohol mixture) to detect certain acid-fast bacilli.

Slide Preparation:
- Prepare a clean, scratch-free slide, immerse it in 95% ethanol, dry it, then heat over a flame.
- Label the slide and mark the smear position on the underside of the slide.
- Collect the clinical specimen (such as sputum, fluid, tissue) or colony from culture, centrifuge to collect the sediment.
- Use a sterile inoculating loop to take one loopful (3mm in diameter) or place one drop of the specimen onto the slideAllow it to air dry and fix by gentle heat
Fixing the Slide: Allow the slide to air dry naturally in a biosafety cabinet or dry it at 60°C.
Staining:
- Flood slide with BacterStain™ Carbon Fuchsin (Code: 04004) and heat 2-3 times until steam rises (Do NOT allow stain to boil or to dry on the slide), let it cool naturally for 5 minutes.
- Wash with water
- Apply several changes of BacterStain™ Alcohol acid (Code: 04009) until no more colour seems to come from the preparation
- Rinse with water and counterstain with BacterStain™ Methylene blue (Code: 04006) for approximately 1 – 2 minutes.
- Decolorize using acid alcohol solution, rinse thoroughly.
- Stain with Methylene blue solution for 1-2 minutes, rinse well.
Rinse with water, drain until dry.
Examine under the microscope, using the oil immersion lens

RESULTS

Mycobacteria: Red/pink color.
Other background fields: Blue color.
AFB Morphology from the specimen: AFB appears as thin, slightly curved rods, staining red, either isolated or clustered together, easily visible against the blue background.

BacterLab ensures the quality of each product batch by testing with ATCC reference strains.

Reference strains	Conditions	Expected results
Positive specimen	oil immersion lens (100x)	AFB stains red on a light blue background, with no staining residue visible
Negative specimen	oil immersion lens (100x)	no AFB is observed

*Note: This ingredient is for reference only our company will formulate the ingredient according to requests or tender specifications.

Product Name	Code	Packaging
BacterStrain™ Ziehl-Neelsen Stain Kit	04002	3 bottles of 100ml/box or according to the customer’s request.