Brucella is a type of non-motile Gram-negative bacterium, with cattle, sheep, pigs and other animals being the most susceptible to infection. Brucella staining is also known as Kozlovsky staining.
Aladdin Brucella Stain Solution uses safranine as the stain. For direct smears of clinical specimens, it yields a clean background, strong contrast between cell nuclei and cytoplasm, and clear and distinguishable intracellular phagosomes, with typical bacterial staining characteristics. All operations should be performed in a Class II biosafety cabinet. If red dot-like bacteria are observed within green-stained bacteria, Brucella infection should be highly suspected. The CDC must be notified promptly to prevent the spread of the epidemic. This reagent is for research use only and is not suitable for clinical diagnosis or other purposes.
Product Components and Storage Conditions:
B1510429
| Component
| 2×50mL
| Storage
|
B1510429A
| Safranine Staining Solution
| 50mL
| RT
|
B1510429B
| Counterstain Solution
| 50mL
| RT |
Materials to Be Prepared:
Inoculation loop or other bacteria-picking tools, alcohol lamp, glass slides, optical microscope
Operating Procedures (For Reference Only):
1. Smear Preparation: Take the bacteria to be tested and spread them into a thin layer in the center of a glass slide; the thinner the better. If a control is required, mix Escherichia coli with the bacteria to be tested and prepare the smear together.
2. Drying: Air-dry the smear naturally at room temperature, or heat it gently over an alcohol lamp to speed up the drying process.
3. Fixation: Hold one end of the glass slide with the specimen side facing up, and move it quickly back and forth 3–5 times over the outer flame of an alcohol lamp, 1 second each time. Do not use excessive heat to avoid denaturation of bacterial proteins. Let the slide cool down before staining.
4. Staining: Add safranine staining solution to cover the smear, and heat it gently over an alcohol lamp until steam appears, which usually takes 30–60 seconds. Rinse the staining solution off with clean water.
5. Counterstaining: Add counterstain solution and stain for 2–3 minutes. Rinse with water, blot dry, and observe under a microscope.
Staining Results:
Brucella
| Red
|
Escherichia coli or other bacteria
| Green
|
Precautions:
1. Mark a circle on the back of the glass slide before preparing the smear to identify the position for subsequent tests.
2. Pay attention to personal protection when sampling bacteria. When removing or replacing the test tube stopper, pass the mouth of the test tube through the flame for brief sterilization. Finally, sterilize the inoculation loop by flaming it thoroughly.
3. When heating the slide for fixation, do not hold the slide too close to the flame. The temperature of the slide should generally not exceed 60°C, which can be judged by touching the back of the slide to the back of your hand—it should feel warm but not scalding.
4. The culture time of the bacteria to be tested can also affect the staining result. Positive bacteria that have been cultured for too long, are already dead, or have undergone lysis often show a negative staining reaction.
5. For your safety and health, wear a lab coat and disposable gloves during the operation.
6. Use the reagent as soon as possible after opening to avoid affecting the results of subsequent experiments.
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