TB Fluorostain Kit is a fluorescent acid-fast staining kit for the detection of Mycobacterium tuberculosis and other acid-fast mycobacteria. The method uses Auramine O-Rhodamine B staining with an Eriochrome Black T counterstain, producing bright orange-yellow fluorescence against a dark background.
The microwave staining procedure takes approximately 10 minutes. Slides can be screened at 100X magnification and examined at 400X for confirmation of mycobacterial morphology. A traditional oven-based staining procedure may also be used.
For in vitro diagnostic use in the United States.
Key Properties
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Product: TB Fluorostain Kit
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Product number: 22422
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Staining method: Auramine O-Rhodamine B fluorochrome staining
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Counterstain: Eriochrome Black T
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Microwave staining time: Approximately 10 minutes
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Screening magnification: 100X
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Confirmation magnification: 400X
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Tests per kit: Approximately 100
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Regulatory use: In vitro diagnostic use in the United States
Applications
Fluorescent Acid-Fast Staining
TB Fluorostain Kit is used for fluorescent staining of acid-fast mycobacteria, including M. tuberculosis. Stained mycobacteria appear bright orange-yellow against a dark background.
Mycobacterial Screening
Fluorochrome-stained slides can be screened at 100X magnification and examined at 400X for confirmation of mycobacterial morphology.
Tissue Sections & Smears
The staining procedure can be used with deparaffinized tissue sections and heat-fixed smears. Preparation guidance is also provided for touch preparations of fresh tissue and body fluids.
Kit Contents
- 500 mL Staining Solution A
- 500 mL Acid Solution B
- 500 mL Counterstaining Solution C
Staining & Preparation Tips
- Use distilled or deionized water throughout the staining procedure; do not use tap water
- Deparaffinize paraffin sections before staining
- Heat-fix smears before staining
- Run a known positive control with each group of slides
- Do not reuse staining solutions between slide groups
- Use a fluorescence microscope with an appropriate filter for Auramine O-Rhodamine B
- For coverslipping, use an aqueous mounting medium such as Aqua-Poly/Mount
Handling & Storage
- Store kit components at room temperature in a cool, well-ventilated location
- Use good laboratory ventilation during staining
- Vent the microwave or operate it in a fume hood when using the microwave procedure
- Keep Acid Solution B away from heat, sparks, open flames, and other ignition sources
- Wear protective gloves, safety glasses, and suitable protective clothing
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FAQ
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1. What is the TB Fluorostain Kit used for?
TB Fluorostain Kit is used for fluorescent staining and detection of Mycobacterium tuberculosis and other acid-fast mycobacteria.
2. What stains are used in the TB Fluorostain Kit?
The method uses Auramine O-Rhodamine B as the fluorescent stain and Eriochrome Black T as the counterstain.
3. How long does TB Fluorostain staining take?
The microwave procedure takes approximately 10 minutes. A traditional oven-based procedure can also be used and requires a little over one hour.
4. What do acid-fast mycobacteria look like after TB Fluorostain staining?
Mycobacteria fluoresce bright orange-yellow against a dark background. Slides can be screened at 100X magnification and examined at 400X for confirmation of morphology.
5. Is TB Fluorostain specific for Mycobacterium tuberculosis?
No. Other acid-fast mycobacteria, including M. leprae and M. avium-intracellulare, can also stain with this fluorochrome method.
6. Can TB Fluorostain slides be restained with carbol-fuchsin?
Yes. Slides stained with TB Fluorostain can be restained using a carbol-fuchsin method for further examination. Slides previously stained with carbol-fuchsin cannot subsequently be restained with the fluorochrome method.
7. How does fluorescent TB staining differ from Ziehl-Neelsen staining?
TB Fluorostain uses an Auramine O-Rhodamine B fluorescence method that can be screened at lower magnification. The AFB Ziehl-Neelsen Kit uses a carbol-fuchsin staining method for examination by standard light microscopy.
8. How many tests are included in the TB Fluorostain Kit?
The kit provides reagents for approximately 100 tests and contains 500 mL each of Staining Solution A, Acid Solution B, and Counterstaining Solution C.
Safety & Documentation
For the complete staining procedure, specimen preparation, microscopy requirements, and interpretation guidance, refer to Technical Data Sheet #488. Review the Safety Data Sheets for Staining Solution A, Acid Solution B, and Counterstaining Solution C before use. Additional staining information is available in the Fluorescent Staining Kits Guide.
