螺旋体 (spirochaeta) 是一类细长、弯曲呈螺旋状、运动活泼的原核细胞型微生物，具有细菌的基本结构，细胞壁有脂多糖和壁酸，螺旋体分为密螺旋体(如梅毒螺旋体)、疏螺旋体(如回归热螺旋体)、钩端螺旋体等；梅毒螺旋体有8～12均匀的螺旋，两端尖直；钩端螺旋体细长，数目较多而且细密，一端或两端弯曲呈钩状，菌体亦常屈曲呈C形或S形。梅毒螺旋体和钩端螺旋体可用镀银法进行显色，如Levaditi法、Warthin-Starry法。
阿拉丁螺旋体染色液 (Levaditi银法) 是利用螺旋体具有的嗜银性，在一定条件下螺旋体可从银液中吸附银离子，经还原液处理后，螺旋体内吸附的银离子被还原为黑色的金属银而显色，主要用于显示梅毒螺旋体，亦可显示钩端螺旋体，但较少显示热回归螺旋体，对肝脏梅毒树胶肿和肝脏钩端螺旋体具有鉴别作用。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：
项目号
| Component
| 3×50mL
| Storage
|
S1510383A
| 银溶液
| 2×50mL
| RT. Store in the dark.
|
S1510383B
| Levaditi 还原液 A
| 25mL
| RT. Store in the dark.
|
S1510383C
| Levaditi 还原液 B
| 25mL
| RT |
临用前，按B:C=1:1混合，即为 Levaditi 还原液，即配即用。
操作步骤(仅供参考)：
1、组织切片厚度1～2mm为佳，置于10%福尔马林中固定2～4天。
2、流水冲洗过夜。
3、95%乙醇浸泡24h，蒸馏水浸洗并不停摇动，直至组织下沉为止。
4、更换新的蒸馏水浸洗10min。
5、入银溶液加盖置于37°C恒温箱孵育1天，更换新的银溶液，继续37°C恒温箱孵育2天。
6、蒸馏水浸洗3次，每次10min。
7、入配制好的Levaditi还原液，加盖室温避光孵育2天。
8、蒸馏水浸洗3次，每次2min。
9、常规脱水透明，浸蜡包埋。
10、切片厚度6～8μm，贴片，烤干。
11、二甲苯或脱蜡透明液脱蜡2次，中性树胶封固。
染色结果：
梅毒螺旋体、钩端螺旋体
| 黑色或棕黑色
|
背景
| 淡黄至棕黄色
|
注意事项：
1、实验所用器皿需用硫酸洗液浸泡，洗干净。
2、步骤4、5的洗片步骤很有必要，一定要把组织中的甲醛和乙醇彻底清除，否则易导致组织内银盐的沉淀。
3、某些细菌和真菌菌丝也会被该染液染成黑色，应注意螺线体为螺旋状形态，并加以鉴别。
4、为了您的安全和健康，请穿实验服并戴一次性手套操作。
5、试剂开封后请尽快使用，以防影响后续实验效果。
Spirochaeta are a class of slender, spiral-shaped, and highly motile prokaryotic microorganisms. They possess the basic structures of bacteria, with lipopolysaccharides and teichoic acids in their cell walls. Spirochaeta are classified into three groups: Treponema (e.g., Treponema pallidum), Borrelia (e.g., Borrelia recurrentis), and Leptospira. Treponema pallidum has 8–12 uniform spirals with pointed and straight ends. Leptospira are slender, with a large number of dense spirals; one or both ends are curved into a hook shape, and the bacterial cells are often bent into a C-shape or S-shape. Treponema pallidum and Leptospira can be visualized by the silver impregnation method, such as the Levaditi method and Warthin-Starry method.
Aladdin Spirochaeta Stain Solution (Levaditi Silver Method) utilizes the argyrophilic property of spirochaeta. Under certain conditions, spirochaeta can adsorb silver ions from the silver solution. After treatment with a reducing solution, the silver ions adsorbed inside the spirochaeta are reduced to black metallic silver, thus achieving color development. This solution is mainly used to visualize Treponema pallidum, and can also be applied to stain Leptospira, but it is less effective for Borrelia recurrentis. It plays a differential role in the identification of hepatic syphilitic gumma and hepatic leptospirosis. This reagent is for research use only and is not suitable for clinical diagnosis or other purposes.
Product Components and Storage Conditions:
S1510383
| Component
| 3×50mL
| Storage
|
S1510383A
| Silver Solution
| 2×50mL
| RT. Store in the dark.
|
S1510383B
| Levaditi Reducing Solution A
| 25mL
| RT. Store in the dark.
|
S1510383C
| Levaditi Reducing Solution B
| 25mL
| RT |
Mix Solution B and Solution C at a ratio of 1:1 to obtain Levaditi Reducing Solution. Prepare immediately before use.
Operating Procedures (For Reference Only):
1. The optimal thickness of tissue sections is 1–2 mm. Fix the sections in 10% formalin for 2–4 days.
2. Rinse the sections under running water overnight.
3. Immerse the sections in 95% ethanol for 24 hours, then rinse them in distilled water with continuous shaking until the tissues sink to the bottom.
4. Replace with fresh distilled water and rinse the sections for 10 minutes.
5. Transfer the sections into silver solution, cover the container, and incubate in a 37°C incubator for 1 day. Replace with fresh silver solution and continue incubating at 37°C for another 2 days.
6. Rinse the sections in distilled water 3 times, 10 minutes each time.
7. Transfer the sections into the prepared Levaditi reducing solution, cover the container, and incubate at room temperature in the dark for 2 days.
8. Rinse the sections in distilled water 3 times, 2 minutes each time.
9. Perform routine dehydration, clearing, paraffin infiltration and embedding.
10. Cut the embedded tissues into sections with a thickness of 6–8 μm, mount the sections onto slides, and dry them by heating.
11. Dewax the sections twice with xylene or dewaxing and clearing solution, then mount the sections with neutral balsam.
Staining Results:
Treponema pallidum, Leptospira
| Black or brownish-black
|
Background
| Pale yellow to yellowish-brown
|
Precautions:
1. All labware used in the experiment must be soaked in sulfuric acid cleaning solution and rinsed thoroughly.
2. The rinsing steps in step 4 and step 5 are essential. Formaldehyde and ethanol in the tissues must be completely removed; otherwise, silver salt precipitation in the tissues will easily occur.
3. Some bacteria and fungal hyphae can also be stained black by this staining solution. Attention should be paid to the spiral morphology of spirochaeta for identification.
4. For your safety and health, please wear a lab coat and disposable gloves during operation.
5. Please use the reagent as soon as possible after opening to avoid affecting the results of subsequent experiments.
ZJ26F0433898
危险品化学品经营许可证（带存储）