Flagella Stain

Flagella Stain

Assay-kits

Product code: ACMBS-58
Specifications: 3x20mL (Box)
Intended use: Intended for staining to observe bacterial whether contain flagella structure
Store temperature: 2℃～8℃
Expiry date: 24 Months

More Details

PrincipleMain componentsSample requirementTest procedureResult interpretationMethod limitationPrecautions

Principle

Flagella is a bacterial organ of movement, must be observed with an electron microscope or use special staining to make flagella thickening and coloring that can be seen under a normal microscope, the staining principle is the latter. Check the bacterial flagella the presence, quantity and location, is one of the commonly methods to identify bacteria. Flagellar composition analysis has great significance for bacterial identification.

Main components

Reagent composition	Main ingredient
1. Carbolfuchsin (solution A)	Carbolfuchsin
2. Tannic acid (solution B)	Tannic acid
3. Potassium alum (solution C)	Potassium alum
4. Filtration bottle	-

Sample requirement

1. Slides processing: Use new slides. First clean slides with cleanser, then dip slides in a 3% hydrochloric acid alcohol over 2 days. Before use, remove the slides from the acid alcohol, rinse with distilled water and air dry or dry with clean gauze.
2. Smear Preparation:

①. First method: Inoculate strains into broth culture tube, incubate them at 37℃ for 24 hours, centrifuge and remove the supernatant. Transfer a loop of precipitate and place it at one end of the slide. Tilt slide to permit bacteria solution run down to the other end of the slide. And dried naturally at room temperature.

②. Second method: Inoculate strains into the bottom of slant agar medium (add some liquid medium or sterile physiological saline at the bottom). After incubating for 8～16 hours, take the colony from the top of slant and suspend in distilled water to obtain a turbid suspension. Remove a loop of the above culture and place it at one end of the slide. Tilt slide to permit suspension to run down to the other end of the slide. And allowed to dry naturally at room temperature.

Test procedure

1. Working Reagent Preparation: Prepare the working reagent by mixing solution A, B and C in a ratio of 1:1:1 and then place it in a filtration bottle. Mix thoroughly and stand for 2 hours prior to use.
2. Add working reagent onto slide and allow staining for about 10～15 minutes.
3. Tilt slide slightly and rinse thoroughly with distilled water drop by drop.
4. Place the slide vertically to permit water to dry out. Air dry and then examine the smear microscopically.

Result interpretation

Both thallus and flagella will appear bright red.

Method limitation

Only use for morphological staining to observe.

Precautions

1. Clean slides are important for flagella staining.
2. After using the working solution, the intensity of the water flushing cannot be too large, otherwise the flagellum and the bacterial body will be easily washed away.
3. This kit must be used by professionals and the interpretation of results also should be made by professionals.
4. Should carefully read the instruction manual before using. Use before expiration date, and keep a good personal hygiene protection.
5. After use, disposal of waste should comply with the hospital or the environmental protection department requirements. Tightly cap the reagent bottle immediately after use to avoid evaporation.
6. Production lot number and expiration date on the package.

References

1. People's Republic of China national health and Family Planning Commission of medical affairs authority. The clinical laboratory operating procedures M. Fourth edition. Beijing: People's Medical Publishing House, 2014.

2. Tongming Qing: Clinical Laboratory Pathogenic Microbiology (M). Higher Education Press.

3. Ni Yu-xing: Microbiology and microbiological testing experimental guidance (M). People's Health Publishing House.

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