Howard Taylor Ricketts (1871–1910), an American pathologist, discovered the unique pathogen causing Rocky Mountain spotted fever in 1909 and later lost his life to this disease. Hence, the pathogen was named Rickettsia. Belonging to Gram-negative bacteria, Rickettsia are a group of Gram-negative prokaryotes that are obligate intracellular parasites of eukaryotic cells. They are intermediate between bacteria and viruses but are more closely related to bacteria, and generally exist in a spherical or rod-shaped form. Rickettsia mainly parasitize arthropods and often cause diseases such as Q fever, Rocky Mountain spotted fever, or typhus. They can be visualized using staining techniques including Gram staining, Giemsa staining, Gimenez staining, and Macchiavello staining.
Gimenez Stain Solution, also known as Gimenez Stain, can stain Rickettsia, Ehrlichia, Wolbachia, viral inclusions, Helicobacter pylori, and other microorganisms. The bacterial cells exhibit colors such as red, purplish red, or dark blue, while the background appears green or teal. Staining results vary depending on the bacterial species. This reagent is intended for research purposes only and is not suitable for clinical diagnosis or any other applications.
Product Components and Storage Conditions:
G1510189
| Component
| 2X50ml
| Storage
|
G1510189A
| Fuchsin Stain Solution
| 16mL
| RT
|
G1510189B
| PB Buffer
| 40 mL
| RT
|
G1510189C
| Malachite Green Stain Solution
| 50 mL
| RT |
Materials to Be Prepared:
Fixatives: 10% neutral formalin, 4% paraformaldehyde, etc.; graded ethanol; distilled water
Operating Procedures (For Reference Only):
I. Staining of Paraffin Sections
1. Fix the tissue in neutral formalin, then perform routine dehydration and embedding.
2. Cut sections with a thickness of 4–6 μm, then deparaffinize and hydrate the sections to water using routine xylene or Deparaffinization and Clearing Solution.
3. Immerse the sections in Gimenez Stain Solution for 2–10 minutes, then rinse with tap water for 30–120 seconds.
4. Immerse the sections in Malachite Green Stain Solution for 10–60 seconds, then rinse with distilled water for 1–5 minutes. Visually confirm that the sections turn blue or blue-green.
5. Blot dry the moisture on the sections and air-dry completely at room temperature.
6. Perform routine dehydration, clear the sections with xylene or deparaffinization and clearing solution, mount with neutral balsam, and observe under an oil immersion lens.
II. Smear Staining
1. Prepare a thin smear on a clean glass slide, air-dry it, and slightly fix the smear by passing it through an alcohol lamp flame.
2. Add Gimenez Stain Solution dropwise and stain for 1–5 minutes, then rinse with tap water for 30–120 seconds.
3. Add Malachite Green Stain Solution dropwise and stain for 10–30 seconds, then rinse with distilled water for 30–180 seconds. Visually confirm that the smear turns blue or blue-green.
4. Blot dry the moisture on the smear and air-dry completely at room temperature.
5. Perform routine dehydration, clear the smear with xylene or deparaffinization and clearing solution, mount with neutral balsam, and observe under an oil immersion lens.
Staining Results:
Rickettsia, Ehrlichia, certain viral inclusions, Helicobacter
| Red–fuchsia–deep purple
|
Wolbachia
| Deep blue
|
Background
| Green–blue–teal |
Precautions:
1. Ensure that the deparaffinization of sections is as thorough as possible.
2. Malachite green staining tends to cause over-staining and uneven staining; please pay close attention to the staining time and water rinsing time.
3. This staining method can effectively demonstrate Legionella in postmortem lung smears.
4. Gimenez Stain Solution can distinguish Rickettsia of different genera: Wolbachia appears deep blue (with a teal background), while Orientia tsutsugamushi appears dark red (other Rickettsia species appear red).
5. Use the reagent as soon as possible after opening to avoid affecting the results of subsequent experiments.
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