Solarbio Flagella Stain Kit (Leifson)

Kit Components

Before use, mix Reagent (A):(B):(C) with a ratio of 1:1:1 to form Leifson Staining Solution. Use it immediately, not for a long time.

Introduction

The bacterial flagellum serves as the motor organ of bacteria. Helicobacter pylori’s ability to navigate through the mucus layer of gastric epithelial cells, from the strongly acidic gastric cavity to the neutral environment of gastric epithelial cells, is a notable example of flagellum movement. Flagella staining allows for the observation of flagella morphology, quantity, and location in bacteria, which are crucial for bacterial identification.

This kit employs the Leifson method, also known as Leifson’s flagella stain method, offering increased sensitivity, simplicity of operation, and reliable results.

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 evenly distribute the bacteria. Avoid grinding or stirring to prevent flagella detachment.
4. Drip Leifson Staining Solution on the slide. When more than half of the area of the Leifson Staining Solution appears metallic gloss film, generally dye for 3-10 minutes. Gently wash the staining solution with distilled water and air-dry naturally.
5. 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 and Flagellum: Red in Different Degrees

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. Operate carefully during the staining process to prevent flagella detachment.
4. Use the prepared Leifson Staining Solution within 2 hours whenever possible to avoid long-term storage.
5. Avoid fixing with high-temperature flame.
6. For safety and health, wear laboratory attire and disposable gloves.