病毒性肝炎是由肝炎病毒引起的肝实质细胞变性坏死为主要病变的传染病，乙型肝炎病毒是世界范围内传播的乙肝病毒，HBsAg存在于肝细胞质内，目前可通过免疫组化方法显示乙型肝炎病毒的HBsAg和HBcAg。显示HBsAg的方法有Shikata地衣红法、醛品红法、维多利亚蓝法等，维多利亚蓝是一种苯甲烷染料，可显示弹力纤维和乙型肝炎表面抗原，不易过染，染色后不易褪色。
阿拉丁乙型肝炎病毒染色液 (Tanaka维多利亚蓝法) 适用于显示弹力纤维和型肝炎表面抗原。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：
H1510443
| Component
| 4×50mL
| Storage
|
H1510443A
| Tanaka 氧化剂 A
| 25mL
| RT. Store in the dark.
|
H1510443B
| Tanaka 氧化剂 B
| 25mL
| RT
|
H1510443C
| Tanaka 漂白剂
| 50mL
| RT
|
H1510443D
| 维多利亚蓝染色液
| 50mL
| RT. Store in the dark.
|
H1510443E
| 核固红染色液
| 50mL
| RT
|
临用前，按A:B=1:1 混合即为Tanaka 氧化剂，不宜提前配制。
自备材料：
1、系列乙醇
2、染色缸
操作步骤(仅供参考)：
1、组织固定于10%中性福尔马林固定液中，常规脱水包埋。
2、切片厚4μm，常规二甲苯或脱蜡透明液脱蜡至水。
3、入Tanaka氧化剂氧化5min，自来水稍洗。
4、入Tanaka漂白剂漂白2min，至上述氧化剂的着色全部脱去。
5、自来水冲洗5min。
6、70%乙醇稍微浸洗。
7、切片入维多利亚蓝染色液的染缸中浸染(加盖)24h。
8、70%乙醇浸洗2次，每次10～30s，期间把切片提起放下，至切片无染色液脱出为止。
9、流水稍冲洗。
10、入核固红染色液复染细胞核5～10min，流水稍冲洗。
11、常规脱水，二甲苯或脱蜡透明液透明，中性树胶封固。
染色结果：
HBsAg 阳性物质
| 蓝色
|
弹力纤维、脂褐素、肥大细胞
| 蓝色
|
细胞核
| 红色
|
注意事项：
1、大多数固定液均可，但含有铬盐的固定液固定后，染色较浅且容易弥散。
2、维多利亚蓝染色液稳定，保存时间长，并可反复使用。
3、维多利亚蓝染色液染色时间在24～48h之间均可，以24h居多。
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。
Viral hepatitis is an infectious disease characterized primarily by degeneration and necrosis of liver parenchymal cells, caused by hepatitis viruses. Hepatitis B virus (HBV) is a hepatitis virus prevalent worldwide, and hepatitis B surface antigen (HBsAg) is present in the cytoplasm of hepatocytes. Currently, immunohistochemical methods can be used to detect HBsAg and hepatitis B core antigen (HBcAg) of HBV. Methods for detecting HBsAg include Shikata’s orcein method, aldehyde-fuchsin method, Victoria blue method, etc. Victoria blue is a type of phenylmethane dye that can stain elastic fibers and hepatitis B surface antigen; it is not prone to over-staining and the stain is resistant to fading after coloring.
Aladdin Hepatitis B Virus Staining Solution (Tanaka Victoria Blue Method) is suitable for demonstrating elastic fibers and hepatitis B surface antigen. This reagent is intended for research use only and is not suitable for clinical diagnosis or any other purposes.
Product Components and Storage Conditions:
H1510443
| Component
| 4×50mL
| Storage
|
H1510443A
| Tanaka Oxidant A
| 25mL
| RT. Store in the dark.
|
H1510443B
| Tanaka Oxidant B
| 25mL
| RT
|
H1510443C
| Tanaka Bleach
| 50mL
| RT
|
H1510443D
| Victoria Blue Staining Solution
| 50mL
| RT. Store in the dark.
|
H1510443E
| Nuclear Fast Red Staining Solution
| 50mL
| RT
|
Prior to use, mix Oxidant A and Oxidant B at a 1:1 ratio to prepare Tanaka Oxidant. Do not prepare the solution in advance.
Materials Required (Self-prepared):
1. Graded ethanol solutions
2. Staining jars
Operating Procedures (For Reference Only):
1. Fix the tissue in 10% neutral formalin fixative, followed by routine dehydration and embedding.
2. Cut into sections with a thickness of 4 μm, then perform routine dewaxing to water using xylene or dewaxing clearing solution.
3. Immerse the sections in Tanaka Oxidant for oxidation for 5 minutes, then rinse briefly with tap water.
4. Transfer the sections into Tanaka Bleach for bleaching for 2 minutes until all the stain from the aforementioned oxidant is completely removed.
5. Rinse the sections with tap water for 5 minutes.
6. Rinse the sections slightly in 70% ethanol.
7. Place the sections in a staining jar containing Victoria Blue Staining Solution, cover the jar, and stain for 24 hours.
8. Rinse the sections twice in 70% ethanol, 10–30 seconds each time. During the rinsing process, lift and lower the sections repeatedly until no more staining solution elutes from the sections.
9. Rinse the sections briefly under running water.
10. Counterstain the cell nuclei with Nuclear Fast Red Staining Solution for 5–10 minutes, then rinse briefly under running water.
11. Perform routine dehydration, clear the sections with xylene or dewaxing clearing solution, and mount the sections with neutral balsam.
Staining Results:
HBsAg-positive Substances
| Blue
|
Elastic Fibers, Lipofuscin, Mast Cells
| Blue
|
Cell Nuclei
| Red
|
Precautions:
1. Most fixatives are applicable; however, fixation with chrome-containing fixatives will result in faint staining with a tendency to diffuse.
2. Victoria Blue Staining Solution is stable, has a long shelf life, and can be reused repeatedly.
3. The staining time with Victoria Blue Staining Solution can range from 24 to 48 hours, with 24 hours being the most commonly adopted duration.
4. For your safety and health, wear a lab coat and disposable gloves during operation.
5. Please use the reagents as soon as possible after opening to avoid affecting subsequent experimental results.
ZJ26F0433836
危险品化学品经营许可证（带存储）