计算溶液所需的质量、体积或浓度。
苏木素伊红 (HE)染色试剂盒 (含分化液和返蓝液)
有货
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。
储存与运输
避光,室温。常规运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
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，是病理学常规制片中最基本的染色方法，应用极其广泛，苏木精是从原产于中南美州的洋苏木中提取出来的浅黄褐色的结晶，是一种碱性染色剂，它在被氧化后生成苏木素，同媒染剂(常用的是三价的铝或盐铁)一起使用，能够使细胞核染色。在病理诊断、教学和科研工作中，常用HE染色对正常组织和病变组织进行形态结构观察，可确定或鉴别病变组织、细胞中出现的某些异常物质与特殊成分，而需要采用的特殊染色方法、酶组织化学方法、免疫组织化学方法等也均是在观察HE染色组织切片的基础上进行的，在HE染色的组织切片中细胞核呈蓝色，细胞浆呈红色，二者形成鲜明的对比，易于观察分析。
苏木素伊红 (HE)染色试剂盒 (含分化液和返蓝液)中苏木素染色液采用自主研发的配方，由进口的高纯度苏木精、氧化剂等组成，不含氧化汞、甲醇等有害物质，对细胞核染色效果好，其特点是不易产生沉淀和金属膜；应用范围广，可以用于人、动物、畜牧、水产等领域，可以用于组织石蜡切片、冰冻切片和组织细胞的染色等。苏木素染色液和伊红染色液均可以重复使用。该产品仅用于科研领域，不宜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。在HE染色中常用0.5-1%盐酸乙醇作为分化液，因酸能破坏苏木素的醌型结构，使组织与色素分离而退色。大多数组织经苏木素染色后，必须用盐酸乙醇分化，使细胞核过多结合的苏木素染料和细胞浆吸附的苏木素染料脱去，再进行伊红染色，才能保证细胞核与细胞浆染色的分明。
4、返蓝作用：分化之后，苏木素在酸性条件下处于红色离子状态，呈红色；在碱性条件下处于蓝色离子状态，呈蓝色。组织切片经酸性乙醇分化后呈红色或粉红色，立即用水除去组织切片上的酸而中止分化，再用弱碱性水使苏木素染上的细胞核呈现蓝色，这个过程称为返蓝作用或蓝化作用，另外用自来水 (尤其是温水)浸洗也可使细胞核返蓝，但所需时间较长。
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自备材料：
1、二甲苯或浸蜡脱蜡透明液、系列乙醇、自来水或蒸馏水
2、乙醚-乙醇混合固定液、4%多聚甲醛、中性树胶或环保封片胶
操作步骤(仅供参考)：
(一)石蜡切片染色
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
①苏木素染色液染色: 3-8min。
②自来水或蒸馏水冲洗: 5-10s。
③酸性乙醇分化: 2-5s。
④自来水冲洗: 20-30s。
⑤返蓝液或温水返蓝: 20-40s。
⑥自来水冲洗: 30-60s。
⑦伊红染色液 (水溶)染色: 20-60s。
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。
(二)冰冻切片染色
1、乙醚-乙醇混合固定液: 5-10s。
2、自来水冲洗: 2-5s。
3、苏木素染色液滴染1-5min (可加热至50℃)。
4、自来水冲洗: 2-5s。
5、酸性乙醇分化: 2-5s。
6、自来水冲洗: 2-5s。
7、返蓝液或温水返蓝: 2-5s。
8、自来水冲洗: 5-10s。
9、伊红染色液 (水溶)染色: 2-20s。
10、自来水冲洗: 5-10s。
11、80%乙醇: 1-2s。
12、95%乙醇: 1-2s。
13、无水乙醇: 2-5s。
14、苯酚二甲苯 (1:3): 2-5s。
15、二甲苯或浸蜡脱蜡透明液透明3次，每次2-5s。
16、中性树胶封片。
(三)细胞染色
1、4%多聚甲醛固定10-20min。
2、自来水冲洗2次，每次2min。
3、蒸馏水冲洗2次，每次2min。
4、染色、脱蜡、透明、封固步骤同石蜡切片的染色步骤，作用时间应相应缩短。
染色结果：
细胞核呈蓝色；
细胞质、肌纤维、胶原纤维、甲状腺胶质等呈深浅不一的红色；
角蛋白、红细胞等呈明亮的橙红色。
注意事项：
1、切片脱蜡应尽量干净；温度低时，可在恒温箱60-70℃处理。
2、系列乙醇应经常更换新液。
3、酸性乙醇分化时间应根据切片厚薄、组织类别以及新旧而定，另外分化后自来水冲洗时间应该足够，以便彻底清洗酸。
4、乙醚-乙醇混合固定液是由乙醚和95%乙醇等量混合而得，再加入适量乙酸，密闭保存。
5、冷冻切片染色时间尽量要短。
6、返蓝液常使用0.2-1%氨水或Scott促蓝液或0.1-1%碳酸锂溶液代替。
7、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, referred to as HE staining for short, is the most basic staining method in routine pathological slide preparation and has extremely wide applications. Hematoxylin is a tawny crystal extracted from logwood, a plant native to Central and South America. It is a basic dye that forms hematein after oxidation. When used together with a mordant (commonly trivalent aluminum or iron salts), it can stain cell nuclei. In pathological diagnosis, teaching, and scientific research, HE staining is commonly used to observe the morphological structure of normal and diseased tissues. It can identify or differentiate certain abnormal substances and special components present in diseased tissues and cells. Special staining methods, enzyme histochemistry, immunohistochemistry, and other techniques are all performed on the basis of observing H&E-stained tissue sections. In H&E-stained sections, cell nuclei stain blue, and cytoplasm stains red, forming a distinct contrast that facilitates observation and analysis.
The Hematoxylin-Eosin (HE) Staining Kit (with Differentiating Solution and Bluing Solution) contains hematoxylin staining solution formulated with an independently developed recipe. The solution is composed of imported high-purity hematoxylin and oxidants, and is free of harmful substances such as mercuric oxide and methanol. It exhibits excellent staining effects on cell nuclei with the characteristics of being less prone to precipitation and metallic film formation. It has a broad application range, suitable for use in human, animal, livestock, and aquatic research fields, and can be applied to the staining of tissue paraffin sections, frozen sections, and tissue cells. Both hematoxylin staining solution and eosin staining solution are reusable.Note: This product is for research use only and not intended for clinical diagnosis or other purposes.
Staining Principles
1. Nuclear Staining Principle: Hematoxylin is a naturally occurring basic dye that can stain 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, making the outer surface of the DNA double helix negatively charged (acidic). This allows it to easily bind to positively charged basic hematoxylin dye via ionic or hydrogen bonds, resulting in staining. Hematoxylin appears blue in alkaline solutions, so cell nuclei are stained blue.
2. Cytoplasmic Staining Principle: Eosin is a synthetic acidic dye that can stain cytoplasm under certain conditions. The main component of cytoplasm is protein, an amphoteric compound. 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 be stained by negatively charged acidic dyes. Eosin dissociates into negatively charged anions in water, which bind to positively charged cations of cytoplasmic proteins, staining the cytoplasm red.
3. Differentiation Principle: After staining, the process of removing excess bound dye from tissues using specific solutions is called differentiation, and the solution used is called a differentiating solution. In HE staining, 0.5-1% hydrochloric acid alcohol is commonly used as the differentiating solution. Acids can destroy the quinone structure of hematoxylin, causing the dye to dissociate from tissues and fade. Most tissues must be differentiated with hydrochloric acid alcohol after hematoxylin staining to remove excess hematoxylin bound to cell nuclei and hematoxylin adsorbed by cytoplasm before eosin staining. This step ensures clear differentiation between nuclear and cytoplasmic staining.
4. Bluing Principle: After differentiation, hematoxylin exists in a red ionic state (appearing red) under acidic conditions and in a blue ionic state (appearing blue) under alkaline conditions. Tissue sections turn red or pink after differentiation with acidic alcohol. Immediately rinse the sections with water to remove acid and terminate differentiation, then treat them with weakly alkaline water to restore the blue color of hematoxylin-stained nuclei. This process is called bluing or regeneration of blue color. Alternatively, rinsing with tap water (especially warm water) can also achieve bluing, but it requires a longer time.
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Materials to Be Prepared by User
1. Xylene or paraffin-embedded deparaffinization and clearing solution, graded ethanol, tap water or distilled water
2. Ether-ethanol mixed fixative, 4% paraformaldehyde, neutral balsam or environment-friendly mounting medium
Operating Procedures (For Reference Only)
(1) Staining of Paraffin Sections
1. Deparaffinization and Hydration of Sections
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 Stain with hematoxylin staining solution: 3-8 minutes.
② Rinse with tap water or distilled water: 5-10 seconds.
③ Differentiate with acid alcohol: 2-5 seconds.
④ Rinse with tap water: 20-30 seconds.
⑤ Bluing with bluing solution or warm water: 20-40 seconds.
⑥ Rinse with tap water: 30-60 seconds.
⑦ Stain with aqueous eosin staining solution: 20-60 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 95% ethanol twice, 1-2 minutes each time.
④ 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 balsam.
(2) Staining of Frozen Sections
1. Fix with ether-ethanol mixed fixative: 5-10 seconds.
2. Rinse with tap water: 2-5 seconds.
3. Stain with hematoxylin staining solution (dropwise application) for 1-5 minutes (heating to 50℃ is optional).
4. Rinse with tap water: 2-5 seconds.
5. Differentiate with acid alcohol: 2-5 seconds.
6. Rinse with tap water: 2-5 seconds.
7. Bluing with bluing solution or warm water: 2-5 seconds.
8. Rinse with tap water: 5-10 seconds.
9. Stain with aqueous eosin staining solution: 2-20 seconds.
10. Rinse with tap water: 5-10 seconds.
11. 80% ethanol: 1-2 seconds.
12. 95% ethanol: 1-2 seconds.
13. Anhydrous ethanol: 2-5 seconds.
14. Phenol-xylene (1:3): 2-5 seconds.
15. Clear with xylene or paraffin-embedded deparaffinization and clearing solution three times, 2-5 seconds each time.
16. Mount with neutral balsam.
(3) Cell Staining
1. Fix with 4% paraformaldehyde for 10-20 minutes.
2. Rinse with tap water twice, 2 minutes each time.
3. Rinse with distilled water twice, 2 minutes each time.
4. Follow the staining, deparaffinization, clearing, and mounting steps for paraffin sections, with appropriately shortened incubation times.
Staining Results
Cell Nuclei: Blue
Cytoplasm, Muscle Fibers, Collagen Fibers, Thyroid Colloid, etc.: Red of varying intensities
Keratin, Red Blood Cells, etc.: Bright Orange-Red
Precautions
1. Ensure thorough deparaffinization of sections. At low temperatures, sections can be treated in an incubator at 60-70℃.
2. Replace graded ethanol with fresh solutions regularly.
3. The duration of acid alcohol differentiation should be adjusted according to section thickness, tissue type, and reagent freshness. In addition, the tap water rinsing time after differentiation should be sufficient to completely remove residual acid.
4. The ether-ethanol mixed fixative is prepared by mixing equal volumes of ether and 95% ethanol, adding an appropriate amount of acetic acid, and storing it in a sealed container.
5. Minimize staining time for frozen sections.
6. Bluing solution can be replaced with 0.2-1% ammonia water, Scott’s bluing solution, or 0.1-1% lithium carbonate solution.
7. For your safety and health, wear a lab coat and disposable gloves during operation.
规格
别名
苏木素伊红 (HE)染色试剂盒
英文别名
Hematoxylin-Eosin (HE) Staining Kit
规格或纯度
BioReagent, 生物染色剂, 用于显微镜
稳定性与储存
各组分在相应的储存条件下保质期1年。
英文名称
Hematoxylin-Eosin (HE) Staining Kit (with Differentiating Solution and Bluing 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
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0737699 |
分析证书 | H1508459 |
| ZJ26F0333019 |
分析证书 | H1508459 |
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