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Staining of bacterial flagella
Description

Hardy Diagnostics Flagella Stain is recommended for use in detecting the presence and arrangement of flagella on the bacterial cell.

SUMMARY

Bacterial flagella, due to their narrow diameter, cannot be seen with a light microscope. The Flagella Stain provides a method for viewing bacterial flagella by employing crystal violet in an alcoholic solution as the primary stain. During the staining procedure, the alcoholic solution evaporates and leaves a precipitate around the flagella, increasing its apparent size to the human eye. The Flagella Stain also contains tannic acid and aluminum potassium phosphate as mordants, and phenol as an antifungal agent.Hardy Diagnostics Flagella Stain is a simple, rapid, qualitative method for detecting bacterial flagella and their shape, length, curvature, arrangement and number on the cell. This method was developed by Ryu in 1937, and also later described by Kodaka, et al. in 1982. This test has been found especially useful in providing taxonomic and identifying information about motile bacteria, and more recently, anaerobic bacteria.

PROCEDURE

Specimen Collection: This product is not intended for primary isolation of patient specimens. It should be used only with cultures of isolated organism. This product used with other biochemical tests to identify cultures of isolated organism. Consult listed references for information on specimen collection.

Organism Preparation:

For best results it is recommended that an 18-48 hour old, well isolated colony taken from a Blood Agar plate be used for the Flagella Stain.Streak the organism to be examined for isolation on a Blood Agar plate (Cat. no. A10). Incubate in an appropriate environment at 35 degrees C. Some non-fermentative organisms grow better at 25 degrees C., and are more likely to be flagellated at this growth temperature.

Smear Preparation:

1. With a wax pencil, draw a border around the clear portion of a microscope slide.

2. Approximately one centimeter from the frosted edge, place a drop of deionized (or distilled) water.

3. With a sterile inoculating loop, gently touch a colony of the culture to be examined and then lightly touch the drop of water without touching the slide. Do not mix.

4. Tilt the slide so the drop will flow to the opposite end of the slide.5. Allow the slide to air dry, at room temperature. Do not heat fix.

Staining Procedure:

1. Flood the slide with Flagella Stain. Let the slide sit for approximately four minutes. However, staining time may need to be adjusted to between two to eight minutes, depending on the age of the stain, air currents and room temperature, and staining solution depth over the smear.

2. Carefully rinse stain by leaving slide on staining rack and allowing water to flow over the surface of the slide. Do not tilt slide while rinsing.

3. Once the stain is washed off, gently tilt the slide to allow excess water to run off.

4. Air dry slide at room temperature.

5. Examine the slide microscopically using the oil immersion objective. Begin the examination at thinner areas of the smear and work towards the center. Look for fields with several isolated cells rather than clumps of bacteria.

INTERPRETATION OF RESULTS

When examined under oil immersion, bacteria and their flagella should stain purple. Consult appropriate references for descriptions, drawings or pictures for specific information.

 

Cat. no. HDZ87

Size: 15mL

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