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BioReagent,生物染色剂,用于显微镜 级 ,适用于对基线干扰要求严格的色谱和分析工作流程。
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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。
阿利新蓝 (Alcian)又称爱先蓝或阿尔辛蓝等,是一种类铜钛花青共轭染料,最初用于纺织纤维染色。这种阳离子染料与酸性基团结合,也即阿尔辛蓝与组织内含有的阴离子基团如羧基和硫酸根形成不溶性复合物。阿利新蓝由中央含铜的酞菁环与四个异硫脲基通过硫醚键相连而成。该异硫脲基呈中度碱性,使阿利新蓝带阳离子。阿利新蓝使碳水化合物着色的确切机制不明,普遍认为是阳离子的异硫脲基通过静电与组织内的多聚阴离子相连,如含羧基和硫酸根的酸性黏液物质的羧基和硫酸根形成不溶性复合物,即染料分子中带正电荷的盐键和酸性黏液物质中带负电荷的酸性基团结合呈蓝色。
黏液HID-AB染色原理在于高铁二胺盐中的二铵盐与硫酸化酸性黏液物质结合,形成复合物而被显色。在pH值大大低于2.5时,组织内的硫酸根电离,带有一个负电荷,与阿利新蓝中的阳离子形成盐键,使带有硫酸根的组织 (如硫酸黏液物质) 染色,硫酸化酸性黏液物质等形成棕紫色至棕黑色。在pH值=2.5 时,组织内的羧基电离,带有一个负电荷,与阿利新蓝中的阳离子形成盐键,使带有羧基的组织 (如蛋白多糖/透明质酸以及上皮酸性黏蛋白) 染色,唾液酸性黏液物质等形成蓝色。黏液HID-AB染色液主要用于鉴别硫酸化酸性黏液物质和唾液酸性粘液物质,小肠上皮产生氮乙酰化诞液酸性黏液物质,大肠上皮产生氧乙酰化诞液酸性黏液物质和硫酸化酸性黏液物质。该法可配合AB-PAS染色,对肠上皮化生的类型进行鉴定,对转移性肿瘤发生黏液的类型进行鉴定,研究胃肠道肿瘤细胞的性质以及协助确定原发肿瘤是否来源于大肠。
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使用方法
1、切片脱蜡至蒸馏水。
2、入配制好的HID Solution浸泡18-24h (20-25℃)。
3、流水冲洗5min。
4、入Alcian染色液,染色10-20min。
5、稍水洗。
6、入核固红染色液,复染10min。
7、流水冲洗1min。
8、梯度乙醇脱水,二甲苯透明,中性树胶封片。
染色结果
硫酸化酸性黏液物质 (如硫酸黏蛋白和唾液黏蛋白): 棕紫色至棕黑色
羧基化蛋白酸性黏液物质 (如蛋白多糖和透明质酸): 蓝色
细胞核: 红色
注意事项
1、固定液采用10%中性福尔马林。
2、HID Solution不宜提前配制。用后可存储于4℃,仍可用1-2次,但特异性不佳。
3、HID Solution对人体有一定损害,请小心操作,避免接触人体皮肤。
4、为了您的安全和健康,请穿实验服并戴一次性手套操作。
Alcian Blue, also known as Alcian or Alician Blue, is a copper phthalocyanine conjugate dye originally used for dyeing textile fibers. As a cationic dye, it binds to acidic groups, specifically forming insoluble complexes with anionic groups (such as carboxyl and sulfate groups) present in tissues. Alcian Blue consists of a central copper-containing phthalocyanine ring linked to four isothiourea groups via thioether bonds. These isothiourea groups exhibit moderate basicity, endowing Alcian Blue with cationic properties. The exact mechanism by which Alcian Blue stains carbohydrates remains unclear, but it is generally believed that the cationic isothiourea groups bind electrostatically to polyanions in tissues—for instance, forming insoluble complexes with carboxyl and sulfate groups in acid mucins containing such groups. In essence, positively charged salt linkages in the dye molecules combine with negatively charged acidic groups in acid mucins, resulting in a blue coloration.
The principle of Mucin HID-AB Staining lies in the reaction where diamine salts in high iron diamine bind to sulfated acid mucins, forming colored complexes. When the pH is well below 2.5, sulfate groups in tissues ionize to carry a negative charge, which then forms salt bonds with cations in Alcian Blue. This process stains tissues containing sulfate groups (e.g., sulfated mucins), rendering sulfated acid mucins in shades ranging from brownish purple to brownish black. At pH 2.5, carboxyl groups in tissues ionize to carry a negative charge, forming salt bonds with cations in Alcian Blue. This stains tissues with carboxyl groups (e.g., proteoglycans/hyaluronic acid and epithelial acid mucins), coloring sialomucins blue. Mucin HID-AB Staining Solution is primarily used to differentiate between sulfated acid mucins and sialomucins. Small intestinal epithelium produces N-acetylated sialomucins, while large intestinal epithelium produces O-acetylated sialomucins and sulfated acid mucins. Combined with AB-PAS staining, this method can identify the type of intestinal metaplasia, determine the type of mucin produced by metastatic tumors, study the properties of gastrointestinal tumor cells, and assist in confirming whether primary tumors originate from the large intestine.
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Operating Procedure
1. Deparaffinize sections and rinse with distilled water.
2. Immerse sections in the prepared HID Solution for 18-24 hours at 20-25℃.
3. Rinse thoroughly with running water for 5 minutes.
4. Transfer sections to Alcian Stain and incubate for 10-20 minutes.
5. Rinse sections briefly with water.
6. Counterstain with Nuclear Fast Red Stain for 10 minutes.
7. Rinse with running water for 1 minute.
8. Dehydrate sections through a graded ethanol series, clear with xylene, and mount with neutral balsam.
Staining Results
Sulfated acid mucins (e.g., sulfomucins and sialomucins): Brownish purple to brownish black
Carboxylated acid mucins (e.g., proteoglycans and hyaluronic acid): Blue
Cell nuclei: Red
Precautions
1. Use 10% neutral buffered formalin as the fixative.
2. HID Solution should not be prepared in advance. After use, it can be stored at 4℃ and reused 1-2 times, but note that the staining specificity may be compromised.
3. HID Solution is harmful to the human body; handle with care and avoid skin contact.
4. For your safety and health, wear a lab coat and disposable gloves during operation.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | M1508677 | |
| 分析证书 | M1508677 | |
| 分析证书 | M1508677 | |
| 分析证书 | M1508677 |
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