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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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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。
甲苯胺蓝(Toluidine Blue O)是一种常用的人工合成染料,属于醌亚胺染料类,这类染料主要含有胺基和醌型苯环两个发色团,从而成色原显色。甲苯胺蓝中的阳离子有染色作用,组织细胞的酸性物质与其中的阳离子相结合而被染色,还含有两个助色团,能促使染料产生电离成盐类,帮助发色团对组织产生染色力,使切片上的组织细胞着色,可染细胞核使之呈蓝色,另外肥大细胞胞质内含有肝素和组织胺等异色性物质,遇到甲苯胺蓝可呈异染性紫红色,临床上经常用于肥大细胞染色。
甲苯胺蓝水溶液 (1%, McIlvaine) 由甲苯胺蓝、磷酸盐、柠檬酸、去离子水等配制而成,一般用于特殊细胞的染色,不推荐用于软骨、胃粘膜等难染组织的染色。该试剂仅用于科研领域,不适用于临床诊断或其他用途。
自备材料:
1、蒸馏水或去离子水、系列乙醇、冰乙酸
2、二甲苯或环保脱蜡透明液、中性树胶
操作步骤(仅供参考):
(一)肥大细胞染色
1、常规二甲苯或脱蜡透明液脱蜡至水。
2、根据切片厚度和组织的不同,浸染于甲苯胺蓝染色液10~30min。
3、蒸馏水或去离子水轻轻冲洗3~5min。
4、(可选)0.5%冰乙酸分化,直到细胞核和颗粒清晰可见。
5、快速95%和无水乙醇脱水,二甲苯或脱蜡透明液透明,中性树胶封固。
染色结果:肥大细胞呈紫红色;背景呈淡蓝色。
(二)细胞涂片染色
1、细胞涂片后,立即放入95%的乙醇中固定15s,取出放在纸巾上。
2、滴加1~2滴稀释后的甲苯胺蓝染色液进行滴染,加盖玻片让染料渗透到细胞中。
3、10~15s后将玻片竖起,稍加压力,使多余染料被纸巾吸去。
4、无需干燥,直接镜检。
染色结果:细胞核、淋巴细胞呈深蓝色;核仁呈紫红色;红细胞呈橘红;细胞质、单核细胞呈淡蓝色。
注意事项:
1、针对于胃粘膜组织、软骨组织等较难着色组织的染色,浸染于甲苯胺蓝染色液的时间应相应延长。
2、为了您的安全和健康,请穿实验服并戴一次性手套操作。
Toluidine Blue O is a commonly used synthetic dye belonging to the quinone-imine dye class. Dyes of this class mainly contain two chromophores, namely amino groups and quinonoid benzene rings, which form chromogens to produce color. The cations in toluidine blue exert the staining effect: acidic substances in tissue cells combine with these cations and thus get stained. The dye also contains two auxochromes that promote its ionization into salts, enhance the staining power of chromophores on tissues, and enable the tissue cells on sections to be colored—specifically, it can stain cell nuclei blue. In addition, the cytoplasm of mast cells contains metachromatic substances such as heparin and histamine, which turn metachromatic purplish red when exposed to toluidine blue. For this reason, toluidine blue is frequently used for mast cell staining in clinical practice.
Toluidine Blue O Aqueous Solution (1%, McIlvaine) is prepared from toluidine blue, phosphate, citric acid, deionized water and other components. It is generally used for the staining of special cell types and is not recommended for staining hard-to-stain tissues such as cartilage and gastric mucosa. This reagent is for research use only and not intended for clinical diagnosis or any other applications.
Materials to be prepared by the user:
1. Distilled water or deionized water, graded ethanol, glacial acetic acid
2. Xylene or environment-friendly deparaffinization and clearing solution, neutral balsam
Operating Procedures (For Reference Only):
(I) Mast Cell Staining
1. Perform routine deparaffinization to water with xylene or deparaffinization and clearing solution.
2. Immerse the sections in toluidine blue staining solution for 10–30 minutes, depending on the section thickness and tissue type.
3. Rinse gently with distilled water or deionized water for 3–5 minutes.
4. (Optional) Differentiate with 0.5% glacial acetic acid until cell nuclei and granules are clearly visible.
5. Dehydrate rapidly with 95% ethanol and absolute ethanol, clear with xylene or deparaffinization and clearing solution, and mount with neutral balsam.
Staining Results: Mast cells appear purplish red; the background appears pale blue.
(II) Cell Smear Staining
1. Immediately after preparing the cell smear, fix it in 95% ethanol for 15 seconds, then take it out and place it on absorbent paper.
2. Add 1–2 drops of diluted toluidine blue staining solution for drop staining, and cover with a coverslip to allow the dye to penetrate the cells.
3. After 10–15 seconds, stand the slide upright and apply slight pressure to absorb excess dye with absorbent paper.
4. Examine under the microscope directly without drying.
Staining Results: Cell nuclei and lymphocytes appear dark blue; nucleoli appear purplish red; red blood cells appear orange red; cytoplasm and monocytes appear pale blue.
Precautions:
1. For staining hard-to-stain tissues such as gastric mucosal tissue and cartilage tissue, the immersion time in toluidine blue staining solution should be appropriately prolonged.
2. For your safety and health, wear a lab coat and disposable gloves during operation.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | M1511107 |
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