计算溶液所需的质量、体积或浓度。
苏木素-伊红 (HE)高清恒定染色试剂盒 (高清恒染)
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)高清恒定染色试剂盒 (高清恒染)由预染液、苏木素染色液、分化液Ⅰ、分化液Ⅱ、返蓝液、伊红染色液组成。其中苏木素染色液采用雷根自研配方，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。本产品可用于HE常规切片染色中显示组织形态结构。本产品特点是着色牢固，长期保存不褪色，1~2500张切片染色效果基本一致，利于HE染色标准化统一化。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。
| H1508526 | Component | 6×100 mL | 6×500 mL | Storage |
| H1508526A | 预染液 | 100 mL | 500 mL | RT. Store in the dark. |
| H1508526B | 苏木素染色液 | 100 mL | 500 mL | RT. Store in the dark. |
| H1508526C | 分化液Ⅰ | 100 mL | 500 mL | RT. |
| H1508526D | 分化液Ⅱ | 100 mL | 500 mL | RT. |
| H1508526E | 返蓝液 | 100 mL | 500 mL | RT. |
| H1508526F | 伊红染色液 | 100 mL | 500 mL | RT. Store in the dark. |
自备材料：
1、乙醚-乙醇混合固定液、4%多聚甲醛、自来水或蒸馏水
2、环保脱蜡液、环保透明液、系列乙醇、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环保脱蜡液脱蜡2次，每次5-10min。
②无水乙醇2次，每次1-3min。
③95%乙醇: 1min。
④90%乙醇: 1min。
⑤80%乙醇: 1min。
⑥自来水或蒸馏水冲洗: 1min。
2、染色
①预染液处理: 30s。
②苏木素染色液染色: 5min。
③自来水或蒸馏水冲洗: 2min。
④分化液Ⅰ分化: 15s。
⑤分化液Ⅱ分化: 5s。
⑥自来水或蒸馏水冲洗: 2min。
⑦返蓝液返蓝: 1min。
⑧自来水或蒸馏水冲洗: 1min。
⑨80%乙醇: 1min。
⑩伊红染色液染色: 10s。
3、脱水、透明、封固
①95%乙醇: 1min。
②无水乙醇2次，每次1min。
③环保透明液透明2次，每次2min。
④中性树胶或环保封片胶封片。
(二)冰冻切片染色
1、-20℃保存的冰冻切片，室温静置复温待组织稍微晾干后，用甲醇固定1min；新鲜的冰冻切片室温静置数分钟，待OCT包埋剂完全晾干以后开始染色，防止脱片。
2、自来水冲洗5s。
3、预染、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。
(三)细胞染色
1、细胞爬片，经4%多聚甲醛固定5-10min。
2、自来水冲洗2次，每次2min。
3、蒸馏水冲洗2次，每次2min。
4、预染、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。
(四)机染
操作步骤同石蜡切片染色。
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、使用前应仔细阅读说明书，并在产品使用期限内使用。
3、系列乙醇应经常更换新液。
4、产品需按照存放温度保存。如果保存温度低于15℃，苏木素染液中可能有明矾结晶析出，一般不影响产品质量，但可能会减少染色切片数量。如果保存温度高于30℃，会加快苏木素的氧化速度，导致染色能力下降，同时伊红染液的挥发程度会加快。
5、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。
6、建议配套使用环保脱蜡液、环保透明液和环保封片胶进行实验。
7、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Hematoxylin and Eosin combined staining, referred to as HE staining, is the most fundamental staining method in routine pathological section preparation and has extremely wide applications. Hematoxylin is a pale yellowish-brown crystalline substance extracted from logwood native to Central and South America. As a basic dye, it is oxidized to form hematein, which, when used together with a mordant (commonly trivalent aluminum or iron salts), 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 enables the identification or differentiation of certain abnormal substances and special components present in diseased tissues and cells. Moreover, special staining methods, enzyme histochemistry, immunohistochemistry, and other techniques are all performed on the basis of observing HE-stained tissue sections. In HE-stained sections, cell nuclei appear blue and cytoplasm appears red, forming a sharp contrast that facilitates observation and analysis.
Hematoxylin-Eosin (HE) High-Definition Consistent Staining Kit (High-Definition Consistent Staining) consists of pre-stain solution, hematoxylin staining solution, differentiating solution Ⅰ, differentiating solution Ⅱ, bluing solution, and eosin staining solution. Among them, the hematoxylin staining solution adopts a proprietary formula independently developed by Leigen, composed of imported high-purity hematoxylin and oxidants, without harmful substances such as mercuric oxide and methanol, and exhibits excellent staining effects on cell nuclei. This product is suitable for displaying tissue morphological structures in routine HE section staining. It features firm dye binding, non-fading after long-term storage, and consistent staining results for 1 to 2500 sections, which is conducive to the standardization and unification of HE staining. 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 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, making the outer surface of the DNA double helix negatively charged (acidic). It readily binds to positively charged basic hematoxylin dye via ionic bonds or hydrogen bonds, thus being stained. Hematoxylin appears blue in alkaline solutions, so cell nuclei are stained blue.
2. Cytoplasmic Staining Principle: Eosin is a synthetic acidic dye that stains cytoplasm under certain conditions. Cytoplasm is mainly composed of proteins, which are amphoteric compounds. Cytoplasmic staining is closely related to the pH value 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, allowing 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 Effect: 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. The acid destroys the quinone structure of hematoxylin, causing the dye to dissociate from the tissue and fade. Most tissues must be differentiated with hydrochloric acid alcohol after hematoxylin staining to remove excess hematoxylin bound to nuclei and hematoxylin adsorbed to cytoplasm before eosin staining, ensuring clear distinction between nuclear and cytoplasmic staining.
4. Bluing Effect: After differentiation, hematoxylin exists in a red ionic state (red color) under acidic conditions and in a blue ionic state (blue color) under alkaline conditions. Tissue sections appear red or pink after differentiation with acid alcohol. The differentiation is immediately terminated by rinsing the sections with water to remove acid, followed by treatment with weakly alkaline water to restore the blue color of hematoxylin-stained nuclei. This process is called bluing or alkalization. Additionally, rinsing with tap water (especially warm water) can also achieve bluing, but it requires a longer time.
| H1508526 | Component | 6×100 mL | 6×500 mL | Storage |
| H1508526A | Pre-stain Solution | 100 mL | 500 mL | RT. Store in the dark. |
| H1508526B | Hematoxylin Staining Solution | 100 mL | 500 mL | RT. Store in the dark. |
| H1508526C | Differentiating Solution Ⅰ | 100 mL | 500 mL | RT. |
| H1508526D | Differentiating Solution Ⅱ | 100 mL | 500 mL | RT. |
| H1508526E | Bluing Solution | 100 mL | 500 mL | RT. |
| H1508526F | Eosin Staining Solution | 100 mL | 500 mL | RT. Store in the dark. |
Materials to Be Prepared by the User
1. Ether-ethanol mixed fixative, 4% paraformaldehyde, tap water or distilled water
2. Environmentally friendly dewaxing solution, environmentally friendly clearing solution, graded ethanol solutions, neutral gum or environmentally friendly mounting medium
Operating Procedures (For Reference Only)
(1) Paraffin Section Staining
1. Dewaxing and Hydration of Sections
① Dewax with environmentally friendly dewaxing solution twice, 5-10 minutes each time.
② Rinse with anhydrous ethanol twice, 1-3 minutes each time.
③ 95% ethanol: 1 minute.
④ 90% ethanol: 1 minute.
⑤ 80% ethanol: 1 minute.
⑥ Rinse with tap water or distilled water: 1 minute.
2. Staining
① Treatment with pre-stain solution: 30 seconds.
② Staining with hematoxylin staining solution: 5 minutes.
③ Rinse with tap water or distilled water: 2 minutes.
④ Differentiation with differentiating solution Ⅰ: 15 seconds.
⑤ Differentiation with differentiating solution Ⅱ: 5 seconds.
⑥ Rinse with tap water or distilled water: 2 minutes.
⑦ Bluing with bluing solution: 1 minute.
⑧ Rinse with tap water or distilled water: 1 minute.
⑨ 80% ethanol: 1 minute.
⑩ Staining with eosin staining solution: 10 seconds.
3. ehydration, Clearing and Mounting
① 95% ethanol: 1 minute.
② Rinse with anhydrous ethanol twice, 1 minute each time.
③ Clear with environmentally friendly clearing solution twice, 2 minutes each time.
④ Mount with neutral gum or environmentally friendly mounting medium.
(2) Frozen Section Staining
1. For frozen sections stored at -20℃, allow them to warm up at room temperature and air-dry slightly, then fix with methanol for 1 minute. For freshly prepared frozen sections, let them stand at room temperature for several minutes until the OCT embedding medium is completely air-dried before staining to prevent section detachment.
2. Rinse with tap water for 5 seconds.
3. Follow the pre-staining, staining, dehydration, clearing and mounting steps for paraffin sections, with appropriately shortened incubation times.
(3) Cell Staining
1. Fix cell slides with 4% paraformaldehyde for 5-10 minutes.
2. Rinse with tap water twice, 2 minutes each time.
3. Rinse with distilled water twice, 2 minutes each time.
4. Follow the pre-staining, staining, dehydration, clearing and mounting steps for paraffin sections, with appropriately shortened incubation times.
(4) Automated Staining
Follow the same operating procedures as paraffin section staining.
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 dewaxing of sections. When the temperature is low, treat sections in an incubator at 60-70℃.
2. Read the instruction manual carefully before use and use the product within its shelf life.
3. Replace graded ethanol solutions with fresh ones regularly.
4. Store the product at the specified temperature. If stored below 15℃, alum crystals may precipitate in the hematoxylin staining solution, which generally does not affect product quality but may reduce the number of sections that can be stained. If stored above 30℃, the oxidation rate of hematoxylin will accelerate, leading to decreased staining ability, and the volatilization of eosin staining solution will also increase.
5. The differentiation time with acid alcohol should be adjusted according to section thickness, tissue type, and reagent freshness. In addition, sufficient rinsing time with tap water after differentiation is required to completely remove residual acid.
6. It is recommended to use the product in conjunction with environmentally friendly dewaxing solution, environmentally friendly clearing solution and environmentally friendly mounting medium for experiments.
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)高清恒定染色试剂盒
英文别名
Hematoxylin-Eosin (HE) High-Definition Consistent 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) High-Definition Consistent Staining Kit (High-Definition Stable Staining)
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 |
|---|---|---|
 ZJ26F0333021 |
分析证书 | H1508526 |
技术文档和文章
Eosinophil Morphological Observation: Staining Basis, Identification Points, and Research Validation
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Morphological Basis, Method Workflow, and Result Interpretation of Urinary Sediment Microscopy
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线虫与虫卵形态学观察方法
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溶液计算器
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