美国病理学家霍华德·泰勒·立克次 (Howard Taylor Ricketts) (1871-1910年) 在1909年发现洛基山斑疹伤寒的独特病原体并被它夺取生命，故名立克次氏体或立克次体 (Rickettsia），立克次氏体属于革兰氏阴性菌，是介于细菌与病毒之间的接近于细菌的一类专性寄生于真核细胞内的G-原核生物，一般呈球状或杆状。立克次氏体主要寄生于节肢动物，常引起Q热、洛基山斑疹热或伤寒，可通过革兰氏、Giemsa、Gimenez、Macchiavello等染色技术显示。
Gimenez染色液也称作吉曼尼兹染色液，可把立克次体、埃立克体、沃尔巴克体、病毒包涵体、幽门螺旋杆菌等染色，菌体呈红色、紫红色、深蓝色等，背景呈绿色或青绿色，不同细菌染色效果不同。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
产品组分及储存条件：
G1510189
| Component
| 2×50ml
| Storage
|
G1510189A
| 品红染色液
| 16mL
| RT
|
G1510189B
| PB Buffer
| 40 mL
| RT
|
G1510189C
| 孔雀绿染色液
| 50 mL
| RT |
自备材料：
固定液：10%中性福尔马林、4%多聚甲醛等、系列乙醇、蒸馏水
操作步骤(仅供参考)：
(一)石蜡切片染色
1、组织固定于中性福尔马林，常规脱水包埋。
2、切片厚度4~6μm，常规二甲苯或脱蜡透明液脱蜡至水。
3、切片入Gimenez染色液浸染2~10min，自来水冲洗30～120s。
4、入孔雀绿染色液浸染10～60s，蒸馏水冲洗1～5min，肉眼观察切片呈蓝色或蓝绿色。
5、吸干切片水分并放于空气中完全干燥。
6、常规脱水，二甲苯或脱蜡透明液透明，中性树胶封固，油镜观察。
(二)涂片染色
1、干净载玻片制备薄涂片，空气干燥，火焰上将涂片轻微固定。
2、滴加Gimenez染色液染1~5min，自来水冲洗30～120s。
3、滴加孔雀绿染色液染10～30s，蒸馏水冲洗30～180s，肉眼观察切片呈蓝色或蓝绿色。
4、吸干切片水分并放于空气中完全干燥。
5、常规脱水，二甲苯或脱蜡透明液透明，中性树胶封固，油镜观察。
染色结果：
立克次体、埃立克体、某些病毒包涵体、螺杆菌
| 红色-紫红色-深紫色
|
沃尔巴克体
| 深蓝色
|
背景
| 绿色-蓝色-蓝绿色 |
注意事项：
1、切片脱蜡应尽量干净。
2、孔雀绿染色容易出现过染等染色不均的现象，请注意染色时间和水洗时间。
3、本染色法可有效地显示尸体肺涂片中的军团菌。
4、Gimenez染色液可以区分不同属的立克次体，沃尔巴克体属呈深蓝色（背景呈青绿色），羌虫病东方体属呈暗红色（其他立克次体呈红色）。
5、试剂开封后请尽快使用，以防影响后续实验效果。
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.
ZJ26F0333153
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