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Delafield苏木素染色液
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。苏木素成熟的方法有自然氧化法、化学氧化法。自然氧化是指暴露于光和空气中，这个过程比较缓慢，约需 3～6 个月不等，但生成物染色能力可维持很长时间。Ehrlich 和Delafield 苏木素就属于这种。
Delafield 苏木素染液非常稳定，成熟后密封可保存 2 年。对核染色质染得很清晰细致，染色时间也稍长。适用于教学和科研上的制片染色，用此染色液对冷冻切片染色则不理想。
染色原理：
1、细胞核染色的原理：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、细胞浆染色的原理：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1%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。另外用自来水浸洗也可使细胞核返蓝，但所需时间较长。
自备材料：
1、盐酸乙醇分化液
2、蓝化液，如稀氨水、碳酸锂溶液等
3、系列乙醇
4、伊红染色液
5、4%多聚甲醛
操作步骤(仅供参考)：
(一)石蜡切片染色
1、切片脱蜡至水
①二甲苯作用2次，每次5-10min。
②(可选)无水乙醇作用2次，每次3-5min。
③95%的乙醇：3-5min。
④90%的乙醇：3-5min。
⑤80%的乙醇：3-5min。
⑥自来水或蒸馏水冲洗1-3min。
2、染色
①Delafield苏木素染色液染色5-8min。
②自来水或蒸馏水冲洗：5-10s。
③(可选)盐酸乙醇分化：2-5s。
④自来水冲洗 ：20-30s。
⑤(可选)蓝化液返蓝：20-40s。
⑥自来水冲洗：30-60s。
⑦伊红染色液染色：0.5-3min。
⑧自来水冲洗：1-5s。
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)细胞染色
①4%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。系列乙醇应经常更换新液。
2、盐酸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。
3、蓝化液常使用 0.2-1%氨水或Scott 促蓝液或 0.1-1%碳酸锂溶液。
4、为了您的安全和健康，请穿实验服并戴一次性手套操作。
5、本产品仅供科研使用，严禁它用。
The combined staining of Hematoxylin and Eosin is commonly referred to as HE staining, which is the most widely used staining method in pathology and histology. Hematoxylin is a basic natural dye that can stain cell nuclei. The main component of chromatin in the nucleus is DNA. In the double-helix structure of DNA, the phosphate groups on the two nucleotide chains face outward, making the outer side of the DNA double helix negatively charged and acidic. This acidic property allows it to easily bind to the positively charged basic hematoxylin dye via ionic bonds or hydrogen bonds, thereby achieving staining. There are two main methods for the maturation of hematoxylin: natural oxidation and chemical oxidation. Natural oxidation refers to the exposure of hematoxylin to light and air; this process is relatively slow, taking approximately 3 to 6 months, but the resulting product maintains its staining ability for a long time. Ehrlich hematoxylin and Delafield hematoxylin are typical examples of hematoxylins matured by this method.
Delafield hematoxylin staining solution is highly stable and can be stored for 2 years in a sealed container after maturation. It stains nuclear chromatin clearly and delicately, though it requires a slightly longer staining time. It is suitable for staining sections used in teaching and scientific research, but it is not ideal for staining frozen sections.
Staining Principles
1. Principle of Nuclear Staining: Hematoxylin is a basic natural dye capable of staining cell nuclei. The main component of chromatin in the nucleus is DNA. In the double-helix structure of DNA, the phosphate groups on the two nucleotide chains are oriented outward, rendering the outer surface of the DNA double helix negatively charged and acidic. This acidity enables easy binding to the positively charged basic hematoxylin dye through ionic bonds or hydrogen bonds, resulting in staining. Hematoxylin appears blue in an alkaline solution, so cell nuclei are stained blue.
2. Principle of Cytoplasmic Staining: Eosin is a synthetic acidic dye that can stain cytoplasm under specific conditions. The main component of cytoplasm is protein, which is an amphoteric compound. The staining of cytoplasm is closely related to the pH value of the staining solution. When the pH value of the staining solution is below the isoelectric point (4.7-5.0) of cytoplasmic proteins, the cytoplasmic proteins undergo basic ionization, making the cytoplasm positively charged. Consequently, the cytoplasm can be stained by the negatively charged acidic dye (eosin). Eosin dissociates in water into negatively charged anions, which bind to the positively charged cations of cytoplasmic proteins, staining the cytoplasm red.
3. Differentiation: After staining, the process of removing excess dye bound to tissues using specific solutions is called differentiation, and the solution used is known as a differentiating solution. In HE staining, 1% hydrochloric acid-ethanol is commonly used as the differentiating solution. Acids can disrupt the quinone structure of hematoxylin, causing the separation of tissue and pigment and thus decolorization. For most tissues stained with hematoxylin, differentiation with 1% hydrochloric acid-ethanol is essential to remove excess hematoxylin bound to cell nuclei and hematoxylin adsorbed by cytoplasm. Only after this step can eosin staining be performed to ensure a clear distinction between the colors of cell nuclei and cytoplasm.
4. Bluing (Re-blushing): 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 ethanol. To stop the differentiation process, the acid on the tissue sections is immediately removed with water. Subsequently, weakly alkaline water is used to make the hematoxylin-stained cell nuclei appear blue—this process is called bluing or re-blushing. Additionally, rinsing with tap water can also achieve nuclear bluing, but it requires a longer time.
Materials to Be Prepared by the User
1. Hydrochloric acid-ethanol differentiating solution
2. Bluing solution (e.g., dilute ammonia water, lithium carbonate solution)
3. A series of ethanol solutions (for dehydration)
4. Eosin staining solution
5. 4% paraformaldehyde solution
Operating Procedures (for Reference Only)
(1) Staining of Paraffin Sections
1. Dewaxing of Sections to Water
① Xylene treatment: 2 times, 5-10 minutes each time.
② (Optional) Absolute ethanol treatment: 2 times, 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: 1-3 minutes.
2. Staining
① Stain with Delafield hematoxylin staining solution: 5-8 minutes.
② Rinse with tap water or distilled water: 5-10 seconds.
③ (Optional) Differentiation with hydrochloric acid-ethanol: 2-5 seconds.
④ Rinse with tap water: 20-30 seconds.
⑤ (Optional) Bluing with bluing solution: 20-40 seconds.
⑥ Rinse with tap water: 30-60 seconds.
⑦ Stain with eosin staining solution: 0.5-3 minutes.
⑧ Rinse with tap water: 1-5 seconds.
3. Dehydration, Clearing, and Mounting
① 80% ethanol: 10-20 seconds.
② 90% ethanol: 10-20 seconds.
③ 95% ethanol treatment: 2 times, 1-2 minutes each time.
④ Absolute ethanol treatment: 2 times, 2-3 minutes each time.
⑤ Clearing with xylene or paraffin-removing clearing solution: 3 times, 2-3 minutes each time.
⑥ Mount the section with neutral balsam.
(2) Cell Staining
① Fixation with 4% paraformaldehyde solution: 10-20 minutes.
② Rinse with tap water: 2 times, 2 minutes each time.
③ Rinse with distilled water: 2 times, 2 minutes each time.
④ Follow the same staining, dewaxing, clearing, and mounting steps as for paraffin sections, but reduce the treatment time accordingly.
Staining Results
Cell nuclei: Blue.
Cytoplasm, muscle fibers, and collagen fibers: Red (with varying shades).
Keratin, red blood cells, etc.: Bright orange-red.
Preautions
1. Ensure thorough dewaxing of sections. Replace the series of ethanol solutions with fresh ones regularly.
2. The differentiation time with hydrochloric acid-ethanol should be adjusted based on section thickness, tissue type, and the freshness of the staining solution. Additionally, the rinsing time with tap water after differentiation should be sufficient to completely remove residual acid.
3. Common bluing solutions include 0.2-1% ammonia water, Scott’s bluing solution, or 0.1-1% lithium carbonate solution.
4. For your safety and health, wear a lab coat and disposable gloves during operation.
5. This product is for scientific research use only. Any other use is strictly prohibited.
规格
英文别名
Delafield Hematoxylin Stain Solution
规格或纯度
BioReagent,用于显微镜,生物染色剂
稳定性与储存
Store at room temperature long term (24 months). Store in the dark.
英文名称
Delafield 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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技术文档和文章
溶液计算器
常见问题
什么是「生物染色剂」？
「生物染色剂」表示该产品经过加工和检测，适用于生物和生化应用。规格可能包括内毒素限值、微生物控制、无菌性、低 DNase/RNase/蛋白酶活性，以及在相关情况下的生物活性验证。
什么是「BioReagent」？
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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。
本产品如何运输？
本产品按常温标准条件运输，无需温控包装。
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