返蓝染色液由碳酸锂和纯化水组成，主要用于调节苏木素的染色，从紫色变为蓝色。可用于生物组织样本切片的手工操作，也适用于自动染色机的自动染色。
检验原理：细胞中的细胞核是由酸性物质组成，它与碱性染料 (苏木素) 的亲和力较强，而细胞浆则相反，它含有碱性物质，和酸性染料 (伊红) 的亲和力较大。因此，细胞或组织切片经返蓝染色液染色后，红细胞呈桔红色，有核细胞核呈紫蓝色，浆呈浅红、浅灰、粉红色等。
样本要求：经适当分化和水洗的组织样本切片。
储存条件：
返蓝染色液应贮存在4-40℃，相对湿度不超过80%，贮存环境要求较暗处，无腐蚀性气体的环境。产品避免冷冻、禁止倒置。有效期：18个月，在有效期内的已开封染色液建议在开封后6个月内使用完，每次用后及时拧紧瓶盖，以免挥发或变质。
使用方法：
1、石蜡切片于二甲苯I、II中各脱蜡5分钟，然后经无水乙醇、95%乙醇、80%乙醇各1分钟，自来水冲洗1分钟；
2、苏木素染液 (Harris) 染5~10分钟，或苏木素染液 (Gill) 染8~15分钟，自来水洗1~2分钟，至片子上无多余染液脱出；
3、苏木素染液 (Harris) 染色后用分化液分化数秒；若用苏木素染液 (Gill) ，染色后则不需分化；
4、用返蓝染色液蓝化30秒，然后水洗1~2分钟；
5、入伊红染液 (水溶性) 染30~60秒，稍洗；若用伊红染液 (醇溶性)，切片应先经80%乙醇；脱水后，再入伊红染液 (醇溶性) 染色10~30秒 (忌水洗)；
6、在95%乙醇I、II中各调色约10秒，时间不宜过长；
7、在无水乙醇I、II中各脱水1~2分钟，二甲苯I、II中各透明1~2分钟；
8、封片胶封片，镜检。
注意事项：
1、本品仅用于体外诊断。
2、本产品用于生物组织样本切片染色。
3、本产品需由具有病理分析基本知识和样本染色操作技能的病理实验室专业试验人员，严格按照产品使用说明使用。
4、试剂开启后应及时使用，应根据染色效果及染色量及时更换染色液。开启后未使用的试剂应盖好瓶盖密封保存，不得使用过期产品。
5、使用者应按照病理分析试验室的生物安全防护原则进行防护，佩戴防护手套、口罩和眼罩。
6、使用后的试剂应按化学物品处理规定进行妥善处理；生物组织样本应按医疗废弃物的有关规定进行处理。
Dako Bluing Buffer consists of lithium carbonate and purified water. It is mainly used to adjust hematoxylin staining and shift the color from purple to blue. It is applicable to manual staining of biological tissue sections as well as automated staining on slide stainers.
Test Principle:
Cell nuclei are composed of acidic substances and exhibit strong affinity for the basic dye hematoxylin, while cytoplasm contains alkaline substances and binds preferentially to the acidic dye eosin. After bluing treatment with bluing staining solution, red blood cells appear orange-red; nucleated cell nuclei turn purplish-blue; cytoplasm shows pale red, light gray or pink hues.
Sample Requirements:
Tissue sections that have undergone proper differentiation and water rinsing.
Storage Conditions:
Store Bluing Staining Solution at 4–40 °C with relative humidity ≤ 80%, in a dim environment free of corrosive gases. Protect the product from freezing and do not invert the bottle. Shelf life: 18 months. Once opened within the shelf life, finish using the solution within 6 months. Tighten the cap immediately after each use to prevent volatilization and deterioration.
Instructions for Use:
1. Deparaffinize paraffin sections in Xylene I and Xylene II for 5 minutes each. Sequentially dehydrate in absolute ethanol, 95% ethanol and 80% ethanol for 1 minute each, then rinse with tap water for 1 minute.
2. Stain with Harris hematoxylin for 5–10 minutes, or Gill hematoxylin for 8–15 minutes. Rinse with tap water for 1–2 minutes until excess stain no longer leaches from slides.
3. Differentiate slides stained with Harris hematoxylin in differentiation solution for several seconds. No differentiation step is required after Gill hematoxylin staining.
4. Bluing in bluing staining solution for 30 seconds, followed by water rinsing for 1–2 minutes.
5. Stain with water-soluble eosin for 30–60 seconds and perform brief rinsing. For alcohol-soluble eosin: dehydrate slides in 80% ethanol first, then stain with alcohol-soluble eosin for 10–30 seconds (DO NOT rinse with water).
6. Color correction in 95% Ethanol I and 95% Ethanol II for around 10 seconds each; avoid over-incubation.
7. Dehydrate in absolute ethanol I and absolute ethanol II for 1–2 minutes each, then clear in Xylene I and Xylene II for 1–2 minutes each.
8. Mount slides with mounting medium and examine under microscope.
Precautions:
1. For in vitro diagnostic use only.
2. Intended for staining biological tissue sections.
3. This product must be handled exclusively by professional laboratory personnel with fundamental knowledge of pathological analysis and slide staining techniques, strictly in accordance with the instructions.
4. Use the reagent promptly after opening. Replace staining solutions regularly based on staining quality and consumption. Seal unused reagent tightly after opening; expired products must be discarded.
5. Operators shall comply with biosafety guidelines for pathology laboratories and wear protective gloves, masks and eye shields.
6. Dispose of spent reagents per chemical waste regulations. Discard biological tissue specimens following medical waste management standards.
ZJ26F0738080
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