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普鲁士蓝染色试剂盒 (核固红法)
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
含铁血黄素 (Hemosiderin) 是一种血红蛋白源性色素，当组织内出血时，血管中逸出的红细胞被巨噬细胞吞噬并由其溶酶体降解，血红蛋白被分解为不含铁的橙色血质和含铁的含铁血黄素。Perls普鲁士蓝反应 (Prussian blue reaction) 又称为含铁血黄素染色，即经过亚铁氰化钾和稀酸处理后可以产生蓝色，常见于吞噬细胞或间质内，其染色原理在于亚铁氰化钾溶液使三价铁离子从蛋白质中被稀盐酸分离出来，三价铁与亚铁氰化钾反应，生成一种不溶解的蓝色化合物即三价铁的亚铁氰化物。
普鲁士蓝染色用于显示局部组织内的各种出血性病变，常见于吞噬细胞内，可以很好地区分含铁血黄素与其他色素。该染色液稳定性好、不易产生沉淀、应用范围广，可以进行复染。该染色液的复染液采用核固红，是最经典、最常用的复染液。
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使用说明：
临用前，取Perls染色液A、Perls染色液B等量混合即为Perls染色液，不宜提前配制。
（一）石蜡切片染色
1、组织固定于10%中性福尔马林，常规脱水包埋。
2、切片厚度4μm，常规脱蜡至水。
3、蒸馏水水洗1min。
4、切片入Perls染色液 (见注意事项2)，浸染15-30min。
5、蒸馏水充分冲洗2-5min。
6、入核固红染色液，淡染细胞核5-10min。
7、蒸馏水冲洗1-5s。
8、常规脱水透明，中性树胶封固。
（二）冰冻切片染色
1、无需脱蜡，直接迅速用蒸馏水冲洗2-3min。
2、染色、封固步骤同石蜡切片的染色步骤，时间可以相应缩短。
（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、封固步骤同石蜡切片的染色步骤，但操作时间应相应延长。
阴性对照：取相同切片脱蜡至水。入5%草酸孵育2-6h，经Perls stain，其余步骤同上。结果为阴性。
注意事项：
1、切片脱蜡应尽量干净。组织固定常采用10%中性福尔马林，经普通福尔马林长期固定后，组织会有损伤。避免使用酸性固定剂，铬酸盐处理也会妨碍铁的保存。
2、整个操作过程中容器要干净，避免用金属铁制品，洗切片和容器时应以蒸馏水冲洗，因普通水内含铁质。Perls染色液染色时，应根据样本情况调整着色时间。
3、所有切片都应使用同一个阳性对照切片，选择适合的对照非常重要。尸检肺组织是一个很好的对照，包含相当数量的铁阳性巨噬细胞 (心衰细胞)。
4、冰冻切片和细胞染色，最好根据具体情况摸索实验条件。
染色结果：
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Hemosiderin is a hemoglobin-derived pigment. When hemorrhage occurs in tissues, red blood cells leaking out from blood vessels are phagocytosed by macrophages and degraded in their lysosomes. Hemoglobin is broken down into iron-free hematoidin and iron-containing hemosiderin. The Perls Prussian blue reaction, also known as hemosiderin staining, produces a blue color upon treatment with potassium ferrocyanide and dilute acid, and positive signals are commonly observed in phagocytes or interstitial tissues. Its staining principle is as follows: dilute hydrochloric acid separates ferric ions from proteins in the presence of potassium ferrocyanide solution. Ferric ions then react with potassium ferrocyanide to form an insoluble blue complex, ferric ferrocyanide.
Prussian blue staining is used to visualize various hemorrhagic lesions in local tissues, which are predominantly located within phagocytes. It enables clear differentiation between hemosiderin and other pigments. This staining solution features excellent stability, low precipitation tendency, a wide range of applicable samples, and compatibility with counterstaining. Nuclear Fast Red is adopted as the counterstain, which is the most classic and widely used counterstaining reagent.
| P774831 | Component | 2×50 mL | 2×100 mL | Storage |
| P774831A | Perls Staining Solution A | 25 mL | 50 mL | RT. |
| P774831B | Perls Staining Solution B | 25 mL | 50 mL | RT. |
| P774831C | Nuclear Fast Red Staining Solution | 50 mL | 100 mL | RT. Store in the dark |
Instructions for Use:
Mix equal volumes of Perls Staining Solution A and Perls Staining Solution B right before use to prepare working Perls staining solution. Pre-mixing in advance is not recommended.
(I) Staining of Paraffin Sections
1. Fix tissues in 10% neutral formalin, followed by routine dehydration and embedding.
2. Cut sections at 4 μm thickness and perform standard deparaffinization to rehydrate with water.
3. Rinse sections with distilled water for 1 min.
4. Immerse sections in Perls working staining solution (see Note 2) for 15–30 min.
5. Thoroughly rinse with distilled water for 2–5 min.
6. Counterstain cell nuclei lightly with Nuclear Fast Red staining solution for 5–10 min.
7. Rinse briefly with distilled water for 1–5 seconds.
8. Conduct routine dehydration and clearing, then mount with neutral balsam.
(II) Staining of Frozen Sections
1. No deparaffinization required; rinse rapidly with distilled water for 2–3 min directly.
2. Follow the same staining and mounting procedures as paraffin sections, with incubation times shortened accordingly.
(III) Cell Staining
1. Fix samples in 4% paraformaldehyde for 10–20 min.
2. Rinse twice with tap water, 2 min each time.
3. Rinse twice with distilled water, 2 min each time.
4. Follow the same staining and mounting procedures as paraffin sections, with incubation times extended appropriately.
Negative Control: Take identical sections and deparaffinize to water. Incubate sections in 5% oxalic acid for 2–6 h, then perform Perls staining with all subsequent steps identical to above. A negative staining result shall be observed.
Precautions:
1. Ensure thorough deparaffinization of sections. Tissues are routinely fixed with 10% neutral buffered formalin; long-term fixation in unbuffered formalin will induce tissue damage. Acidic fixatives and chromate treatments should be avoided, as they impair iron preservation in specimens.
2. All vessels used during the entire staining procedure must be clean. Iron-containing metal tools are forbidden. Only distilled water can be used for rinsing sections and containers, since tap water contains iron impurities. Adjust the Perls staining incubation time according to sample types.
3. The identical positive control section shall be adopted for all batches of sections, and proper control selection is crucial. Autopsy lung tissue is an ideal positive control sample, which contains abundant iron-positive macrophages (heart failure cells).
4. Experimental parameters for frozen sections and cell staining are recommended to be optimized according to practical sample conditions.
Staining Results:
| Hemosiderin / Ferric iron | Blue |
| Cell nuclei and other tissue components | Red |
规格
别名
普鲁士蓝-核固红染色试剂盒 | 铁染色试剂盒 | 普鲁士蓝染色试剂盒 (铁染色)
英文别名
Prussian Blue and Nuclear Fast Red Staining Kit | Iron Stain Kit | Prussian Blue Iron Stain Kit | Perls' Prussian Blue Stain Kit
规格或纯度
BioReagent, 生物染色剂, 用于显微镜
英文名称
Prussian Blue Staining Kit (Nuclear Fast Red)
敏感性
Light-sensitive
储存与运输
储存条件
避光,室温
运输条件
常规运输
稳定性与储存
Store at room temperature long term (12 months). Store in the dark.
技术文档
高级数据
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0737644 |
分析证书 | P774831 |
| ZJ26F0737643 |
分析证书 | P774831 |
| ZJ25F0521427 |
分析证书 | P774831 |
技术文档和文章
生物染色剂
目录： 阿拉丁试剂的规格、级别和纯度
Histological and Cytological Section Staining: Principles and Methodological Essentials for Routine Pathology Stains and Structural Special Stains
目录： 技术文章
Principles, Classification, and Clinical Laboratory Applications of Hematologic Cytochemical Staining
目录： 技术文章
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