Solarbio Flagella Stain Kit (silver stain)

Kit Components

Mix A1 and A2 with a ratio of 1:1 to form Tannic Acid Working Solution before use.

Introduction

• Role of Bacterial Flagellum:
  • The bacterial flagellum acts as the motor organ, facilitating movement in bacteria.
  • Notably, Helicobacter pylori utilizes its flagella to navigate through the mucus layer of gastric epithelial cells, from the highly acidic gastric cavity to the neutral environment of gastric epithelial cells.
• Flagella Staining Importance:
  • Flagella staining allows for the observation of flagella morphology, quantity, and location in bacteria.
  • These observations are crucial for bacterial identification and understanding their motility.
• Kit Description:
  • This kit utilizes the silver staining method for flagella staining.
  • It employs Ammonia Silver Staining Solution, known for its sensitivity and simplicity.
  • The use of this staining solution enhances the reliability of outcomes, providing accurate flagella visualization.

Self Provided Materials

Alcohol lamp, slide, distilled water, inoculation ring, microscope.

Protocol (For reference only)

1. Place 2 drops of distilled water on a clean, grease-free slide.
2. Using the inoculation ring, transfer sterile distilled water onto the slide, then collect bacteria from a blood plate colony, allowing them to swim into the distilled water within the ring. Repeat this process twice.
3. Gently shake the slide to disperse the bacteria. Avoid grinding or stirring to prevent flagella detachment.
4. Air-dry the slide at room temperature or in a 35°C incubator (avoid flame fixation).
5. Heat the Tannic Acid Working Solution in a clean container or test tube using an alcohol lamp. Cool slightly and use immediately.
6. Apply the heated Tannic Acid Working Solution to the slide and stain for 30-40 seconds. Slowly rinse with distilled water and air-dry by shaking.
7. Apply Ammonia Silver Staining Solution, heat gently, rinse with distilled water, and air-dry naturally.
8. Begin microscopic examination from the edge of the smear, gradually moving inward. Flagella are easier to observe where bacteria are less concentrated; they may be obscured by bacterial concentration.

Result

• Thalli: Dark brown
• Flagellum: Brown

Note

1. Ensure slides are clean and free from oil contamination.
2. Bacterial samples for staining must be in the active growth stage, preferably from successive subcultures.
3. Handle with care during staining to prevent flagella detachment. Avoid high-temperature flame fixation.
4. Use the prepared Tannic Acid Working Solution within 10 minutes whenever possible.
5. Ammonia Silver Staining Solution is unstable; store it strictly avoiding light at 4°C and discard if serious turbidity occurs.
6. For safety and health, wear laboratory attire and disposable gloves.