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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
淀粉样物质是一种无固定形状的细胞外嗜酸性物质,可存在于不同的组织、器官,导致的疾病称为淀粉样变;淀粉样物质主要是由蛋白质构成,该蛋白大部分排列成反向的β-折叠层结构。在电子显微镜下淀粉样物质呈原纤维排列,病例材料中为大量细胞外的不分支的细丝,大多随机排列,用于识别淀粉样物质的组织学方法有甲紫染色、刚果红染色、偏振光显微镜观察等。目前研究发现传统的甲紫染色法灵敏度低、特异性差,经典的而且有效的方法是刚果红染色,1922年Bennhold发现了刚果红可以用于活体内淀粉样物质的鉴别,并应用到组织切片。
淀粉样物质染色液 (Puchtler碱性刚果红法)主要由刚果红染色液、苏木素染色液等组成,其染色原理在于淀粉样物质对刚果红比其他的组织结构具有更大的亲和力,其羟基与刚果红的胺基结合,从而使淀粉样物质染成红色,该染色法性能稳定,是非常经典的淀粉样物质染色的方法。该试剂仅用于科研领域,不适用于临床诊断或其他用途。
| A1508466 | Component | 4×50 mL | Storage |
| A1508466A | 苏木素染色液 | 50 mL | RT. Store in the dark. |
| A1508466B | 酸性乙醇分化液 | 50 mL | RT |
| A1508466C | 氯化钠溶液 | 50 mL | RT |
| A1508466D | 刚果红染色液 | 50 mL | RT. Store in the dark. |
| A1508466E | Puchtler碱化液 | 1 mL | RT |
自备材料:
1、10%中性福尔马林、蒸馏水、系列乙醇
2、二甲苯或环保浸蜡脱蜡透明液、中性树胶、Scott蓝化液
操作步骤(仅供参考):
1、常规固定,固定液没有特殊要求,常采用10%的中性福尔马林固定液,常规脱水包埋。
2、切片厚度4μm,常规二甲苯或脱蜡透明液脱蜡至水。
3、入苏木素染色液,浸染5min。
4、酸性乙醇分化2-5s,立即入水终止分化,水洗2次后镜下控制至恰当程度。
5、自来水冲洗2min。
6、入Scott蓝化液或水洗返蓝,自来水冲洗2min。
7、按氯化钠溶液: Puchtler碱化液=100: 1的比例配制碱性氯化钠溶液,即配即用;切片入碱性氯化钠溶液浸染20min。
8、按刚果红染色液: Puchtler碱化液=100: 1的比例配制碱性刚果红染色液,即配即用;切片直接入碱性刚果红染色液浸染20min。
9、无水乙醇轻轻冲洗,二甲苯或脱蜡透明液透明,中性树胶封固。
染色结果:
淀粉样物质: 红色
细胞核: 蓝色
注意事项:
1、切片脱蜡应尽量干净,否则影响染色效果。
2、酸性乙醇分化液应密闭保存,一旦开启尽快用完。
3、刚果红染色液染色时尽量采用浸染,如果滴染,应置于湿盒防止溶液挥发。。
4、碱性氯化钠溶液和碱性刚果红染色液即配即用,尽量在20min内使用。
5、为了您的安全和健康,请穿实验服并戴一次性手套操作。
6、试剂开封后请尽快使用,以防影响后续实验效果。
Amyloid is an amorphous extracellular eosinophilic substance that can be present in various tissues and organs, and the diseases caused by it are referred to as amyloidosis. Amyloid is mainly composed of proteins, most of which are arranged in an antiparallel β-pleated sheet structure. Under an electron microscope, amyloid exhibits a fibrillary arrangement, appearing as a large number of unbranched extracellular filaments in pathological specimens, mostly arranged randomly. Histological methods for identifying amyloid include methyl violet staining, Congo red staining, and polarized light microscopy. Current research has demonstrated that the traditional methyl violet staining method has low sensitivity and poor specificity, while Congo red staining is a classic and effective approach. In 1922, Bennhold discovered that Congo red could be used for the identification of amyloid in vivo and applied this technique to tissue sections.
Amyloid Staining Solution (Puchtler Alkaline Congo Red Method) is mainly composed of Congo Red Staining Solution and Hematoxylin Staining Solution. Its staining principle lies in that amyloid has a higher affinity for Congo red than other tissue structures, and its hydroxyl groups bind to the amino groups of Congo red, thereby staining amyloid red. This staining method has stable performance and is a very classic technique for amyloid staining. This reagent is for research use only and is not suitable for clinical diagnosis or any other purposes.
| A1508466 | Component | 4×50 mL | Storage |
| A1508466A | Hematoxylin Staining Solution | 50 mL | RT. Store in the dark. |
| A1508466B | Acid Ethanol Differentiating Solution | 50 mL | RT |
| A1508466C | Sodium Chloride Solution | 50 mL | RT |
| A1508466D | Congo Red Staining Solution | 50 mL | RT. Store in the dark. |
| A1508466E | Puchtler Alkalizing Solution | 1 mL | RT |
Materials to be Prepared by User
1. 10% neutral formalin, distilled water, graded ethanol
2. Xylene or environment-friendly paraffin removal and clearing solution, neutral balsam, Scott's Bluing Solution
Procedure (for reference only)
1. Perform routine fixation. There is no special requirement for the fixative; 10% neutral formalin fixative is commonly used, followed by routine dehydration and embedding.
2. Cut sections with a thickness of 4 μm, and perform routine dewaxing to water using xylene or dewaxing and clearing solution.
3. Immerse the sections in Hematoxylin Staining Solution for 5 min.
4. Differentiate with Acid Ethanol Differentiating Solution for 2-5 s, immediately transfer the sections to water to terminate differentiation, wash twice with water, and check under a microscope to ensure appropriate differentiation.
5. Rinse with tap water for 2 min.
6. Immerse the sections in Scott's Bluing Solution or rinse with water for bluing, then rinse with tap water for 2 min.
7. Prepare alkaline sodium chloride solution at a ratio of sodium chloride solution: Puchtler alkalizing solution=100:1. Prepare it immediately before use. Immerse the sections in the alkaline sodium chloride solution for 20 min.
8. Prepare alkaline Congo red staining solution at a ratio of Congo red staining solution: Puchtler alkalizing solution=100:1. Prepare it immediately before use. Directly immerse the sections in the alkaline Congo red staining solution for 20 min.
9. Rinse gently with anhydrous ethanol, clear the sections with xylene or dewaxing and clearing solution, and mount them with neutral balsam.
Staining Results
Amyloid: Red
Cell Nucleus: Blue
Precautions
1. Ensure thorough dewaxing of sections; otherwise, the staining effect will be affected.
2. Store Acid Ethanol Differentiating Solution in a sealed container and use it as soon as possible after opening.
3. Prefer immersion staining when using Congo Red Staining Solution; if drop staining is adopted, place the sections in a moist chamber to prevent solution evaporation.
4. Prepare alkaline sodium chloride solution and alkaline Congo red staining solution immediately before use, and try to use them within 20 min.
5. For your safety and health, wear a lab coat and disposable gloves during operation.
6. Use the reagent as soon as possible after opening to avoid affecting the results of subsequent experiments.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | A1508466 |
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