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Heidenhain铁苏木素染色液
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为什么选择此级别
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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避光,室温。常规运输 。请查阅批次 COA 获取详细规格。
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文献证明
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。
概述
苏木素 (Hematoxylin)和伊红 (Eosin)联合染色简称HE染色，是病理学和组织学最常用的一种染色方法。苏木精为碱性天然染料，可使细胞核着色，细胞核内染色质的主要成分是DNA，在DNA的双螺旋结构中，两条核苷酸链上的磷酸基向外，使DNA双螺旋的外侧带负电荷，呈酸性，很容易与带正电荷的苏木精碱性染料以离子键或氢键结合而被染色。
Heidenhain铁苏木素染色液以硫酸铁铵作为氧化剂和分化剂，根据不同的分化程度可显示不同的结构，染色后所有成分均为黑色或深灰黑色，不同组织结构的苏木素着色可被Heidenhain分化液以不同的速度进行性褪去，黑色褪去顺序依次为：线粒体、横纹肌、核染色质。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
染色原理：
1、细胞核染色原理：苏木素为碱性天然染料，可使细胞核着色，细胞核内染色质的成分主要是DNA，在DNA双螺旋结构中两条核苷酸链上的磷酸基向外，使DNA双螺旋的外侧带负电荷，呈酸性，很容易与带正电荷的苏木素碱性染料以离子键或氢键结合而被染色，苏木素在碱性溶液中呈蓝色，所以细胞核被染成蓝色。
2、细胞浆染色原理：伊红是一种化学合成的酸性染料，在一定条件下可使细胞浆着色，细胞浆的主要成分是蛋白质，为两性化合物，细胞浆的染色与染液的pH值密切相关，当染色液pH值在胞浆蛋白质等电点 (4.7-5.0)以下时，胞浆蛋白质以碱式电离，则细胞浆带正电荷，就可被带负电荷的酸性染料染色。伊红在水中离解成带负电荷的阴离子，与胞浆蛋白质带正电荷的阳离子结合，使细胞浆着色，呈现红色。
3、分化作用：染色后，用某些特定的溶液将组织过多结合的染色剂脱去，这个过程称为分化作用，所用的溶液称为分化液，HE染色中常用0.5-1%盐酸乙醇作为分化液。
| H1508463 | Component | 4×100 mL | Storage |
| H1508463A | Heidenhain Differentiation | 2×100 mL | RT |
| H1508463B | Heidenhain铁苏木素染色液 | 100 mL | RT. Store in the dark. |
| H1508463C | 伊红复染液 (水溶)(备选) | 100 mL | RT. Store in the dark. |
自备材料：
1、自来水或蒸馏水、二甲苯或浸蜡脱蜡透明液、盐酸乙醇分化液、系列乙醇、中性树胶
2、蓝化液 (稀氨水、碳酸锂溶液等)、乙醚-乙醇混合固定液、4%多聚甲醛
操作步骤(仅供参考)：
1、切片脱蜡至水
①二甲苯或浸蜡脱蜡透明液作用2次，每次5-10min。
②(可选)无水乙醇作用2次，每次3-5min。
③95%乙醇: 3-5min
④90%乙醇: 3-5min
⑤80%乙醇: 3-5min
⑥自来水或蒸馏水(亦可用30-40℃温水)冲洗: 1-3min
2、染色
①Heidenhain Differentiation媒染: 1h (见注意事项1)
②自来水或蒸馏水冲洗: 5-10s
③Heidenhain铁苏木素染色液染色: 1-2h
④自来水冲洗: 20-40s
⑤用蒸馏水1:1稀释Heidenhain Differentiation分化，并与自来水冲洗交替进行，显微镜下观察分化程度 (见注意事项2)。
⑥自来水冲洗: 20-40min
⑦(可选步骤)伊红染色: 20-120s
⑧自来水冲洗: 30-60s
3、脱水、透明、封固
①80%乙醇: 10-20s
②90%乙醇: 10-20s
③95%乙醇作用2次，每次1-2min。
④无水乙醇作用2次，每次2-3min。
⑤二甲苯或浸蜡脱蜡透明液透明3次，每次2-3min。
⑥中性树胶封片、镜检。
染色结果：线粒体、横纹肌、髓磷脂、染色质等呈灰黑色。
注意事项：
1、Heidenhain Differentiation媒染时间和Heidenhain铁苏木素染色液染色时间根据不同的固定液而异；一般情况下媒染和染色时间控制在1h即可，参考时间为：福尔马林、Bouin固定液、Carnoy固定液1h，Helly、Zenker等重铬酸盐固定液3h，四氧化锇、Flemming固定液24h。
2、显微镜下控制分化程度，直到出现所需观察的结构；若分化过度，可用苏木素重染相同时间并重新分化；亦可用蒸馏水2:1稀释Heidenhain Differentiation后再进行分化，以便更好控制分化程度。
3、切片分化后应彻底冲洗洗掉所有分化液，组织不易褪色。
4、胞浆复染 (伊红或橙黄G)可突出核染色质，尤其在显示染色体或有丝分裂更有效，本试剂提供了伊红复染液，但不是必需步骤。
5、切片脱蜡应尽量干净。
6、系列乙醇应经常更换新液。
7、冷冻切片染色时间尽量要短。
8、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Combined staining with Hematoxylin and Eosin, abbreviated as HE staining, is the most commonly used staining method in pathology and histology. Hematoxylin is a natural basic dye that stains cell nuclei. The main component of chromatin in the nucleus is DNA. In the DNA double helix structure, the phosphate groups on the two nucleotide chains face outward, rendering the outer side of the DNA double helix negatively charged and acidic. This allows it to readily bind to the positively charged basic dye hematoxylin via ionic or hydrogen bonds, resulting in nuclear staining.
Heidenhain's Iron Hematoxylin Staining Solution uses ammonium ferric sulfate as both an oxidizing agent and a differentiating agent. Different structures can be visualized by controlling the degree of differentiation. After staining, all targeted components appear black or dark grayish-black. The hematoxylin staining of different tissue structures can be progressively decolorized by Heidenhain's differentiating solution at varying rates. The order of decolorization is as follows: mitochondria → striated muscle → nuclear chromatin.This reagent is for research use only and not intended for clinical diagnosis or other applications.
Staining Principles
1. Nuclear Staining PrincipleHematoxylin is a natural basic dye that stains cell nuclei. Chromatin in the nucleus is primarily composed of DNA. In the DNA double helix, phosphate groups on the two nucleotide chains face outward, giving the helix a negatively charged, acidic outer surface. This acidic region easily binds to the positively charged basic dye hematoxylin through ionic or hydrogen bonds. Hematoxylin appears blue in alkaline solutions, thus staining cell nuclei blue.
2. Cytoplasmic Staining PrincipleEosin is a synthetic acidic dye that stains cytoplasm under appropriate conditions. Cytoplasm is mainly composed of proteins, which are amphoteric compounds. Cytoplasmic staining is closely related to the pH of the staining solution. When the pH of the staining solution is below the isoelectric point of cytoplasmic proteins (4.7-5.0), cytoplasmic proteins undergo basic ionization and carry a positive charge, enabling them to bind to negatively charged acidic dyes. Eosin dissociates into negatively charged anions in water, which bind to the positively charged cations of cytoplasmic proteins, resulting in red cytoplasmic staining.
3. Differentiation PrincipleAfter staining, the process of removing excess bound dye from tissues using specific solutions is called differentiation, and the solution used is referred to as a differentiating solution. In HE staining, 0.5-1% hydrochloric acid ethanol is commonly used as the differentiating solution.
| H1508463 | Component | 4×100 mL | Storage |
| H1508463A | Heidenhain Differentiation | 2×100 mL | RT |
| H1508463B | Heidenhain Iron Hematoxylin Staining Solution | 100 mL | RT. Store in the dark. |
| H1508463C | Eosin Counterstain Solution (Aqueous, Optional) | 100 mL | RT. Store in the dark. |
Materials to Be Prepared by the User
1. Tap water or distilled water; xylene or paraffin-embedded deparaffinization and clearing solution; hydrochloric acid ethanol differentiating solution; graded ethanol series; neutral mounting medium.
2. Bluing solution (e.g., dilute ammonia water, lithium carbonate solution); ether-ethanol mixed fixative; 4% paraformaldehyde.
Operating Steps (For Reference Only)
1. Deparaffinization of Sections to Water
① Treat sections with xylene or paraffin-embedded deparaffinization and clearing solution twice, 5-10 minutes each time.
② (Optional) Treat with anhydrous ethanol twice, 3-5 minutes each time.
③ 95% ethanol: 3-5 minutes.
④ 90% ethanol: 3-5 minutes.
⑤ 80% ethanol: 3-5 minutes.
⑥ Rinse with tap water or distilled water (30-40℃ warm water is also acceptable): 1-3 minutes.
2. Staining
① Mordant with Heidenhain's Differentiating Solution: 1 hour (see Precautions 1).
② Rinse with tap water or distilled water: 5-10 seconds.
③ Stain with Heidenhain's Iron Hematoxylin Stain Solution: 1-2 hours.
④ Rinse with tap water: 20-40 seconds.
⑤ Dilute Heidenhain's Differentiating Solution with distilled water at a 1:1 ratio for differentiation. Alternate differentiation with tap water rinses, and observe the degree of differentiation under a microscope (see Precautions 2).
⑥ Rinse with tap water: 20-40 minutes.
⑦ (Optional step) Eosin staining: 20-120 seconds.
⑧ Rinse with tap water: 30-60 seconds.
3. Dehydration, Clearing, and Mounting
① 80% ethanol: 10-20 seconds.
② 90% ethanol: 10-20 seconds.
③ Treat with 95% ethanol twice, 1-2 minutes each time.
④ Treat with anhydrous ethanol twice, 2-3 minutes each time.
⑤ Clear with xylene or paraffin-embedded deparaffinization and clearing solution three times, 2-3 minutes each time.
⑥ Mount with neutral mounting medium and observe under a microscope.
Staining Results
Mitochondria, striated muscle, myelin, chromatin, and other targeted structures appear grayish-black.
Precautions
1. The mordanting time with Heidenhain's Differentiating Solution and staining time with Heidenhain's Iron Hematoxylin Staining Solution vary depending on the fixative used. Generally, both mordanting and staining can be controlled at 1 hour. Reference times are as follows: 1 hour for formalin, Bouin's fixative, and Carnoy's fixative; 3 hours for dichromate fixatives such as Helly's and Zenker's fixatives; 24 hours for osmium tetroxide and Flemming's fixative.
2. Control the degree of differentiation under a microscope until the desired structures are clearly visualized. If over-differentiation occurs, restain with hematoxylin for the same duration and repeat differentiation. Alternatively, dilute Heidenhain's Differentiating Solution with distilled water at a 2:1 ratio to facilitate better control of the differentiation process.
3. After differentiation, thoroughly rinse sections to remove all residual differentiating solution, which prevents unintended decolorization of tissues.
4. Cytoplasmic counterstaining (with eosin or orange G) enhances the contrast of nuclear chromatin, and is particularly effective for visualizing chromosomes or mitotic figures. Eosin counterstain solution is provided with this reagent, but it is not a mandatory step.
5. Ensure thorough deparaffinization of sections.
6. Replace the graded ethanol series with fresh solutions regularly.
7. Minimize staining time for frozen sections.
8. For your safety and health, wear a lab coat and disposable gloves during operation.
规格
英文别名
Heidenhain's Iron Hematoxylin Staining Solution
规格或纯度
BioReagent,用于显微镜,生物染色剂
稳定性与储存
各组分在相应的储存条件下保质期2年。
英文名称
Heidenhain Iron Hematoxylin Staining Solution
储存条件
避光,室温
运输条件
常规运输
名称和识别符
分子类型
生物试剂/缓冲液
技术文档
高级数据
化学和物理性质
敏感性
Light-sensitive
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0333267 |
分析证书 | H1508463 |
技术文档和文章
溶液计算器
常见问题
什么是「生物染色剂」？
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本产品应如何储存？
请于室温、避光条件下保存。本品对光敏感，请保存在原始的避光或棕色容器中。
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