计算溶液所需的质量、体积或浓度。
致畸试验骨组织染色液 (茜素红贮备液)
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概述
致畸试验骨组织染色液 (茜素红贮备液) 由茜素红、冰乙酸、甘油、水合氯醛按国标配比复配稳定剂制成，外观为棕色澄清液体，存在轻微刺激性。适用于食品、化妆品、保健用品、工业化学品毒理学安全性评价，适用于大鼠、小鼠、家兔致畸试验胎仔骨骼形态学检查。
染色原理：茜素红可特异性与骨组织钙盐螯合生成稳定紫红色复合物，完整标记胎仔全身骨化中心、骨缝、肋骨、脊柱、四肢骨；经KOH水溶液稀释后适配致畸试验胎仔整体透明染色流程，可清晰识别骨缺损、骨骼融合、肋骨分叉、骨化不全等致畸相关骨骼发育畸形。
该试剂仅用于科研领域，不适用于临床诊断或其他用途。
操作步骤(仅供参考)：
一、工作液(茜素红应用液)配制
取本品致畸试验骨组织染色液 (茜素红贮备液) (B1529932) 10mL，加入10-20g/L氢氧化钾(KOH)水溶液定容至1000mL，充分摇匀。
注：配制操作需在通风环境中进行，佩戴防护手套、护目镜。应用液现配现用，装入棕色瓶避光存放，3日内使用完毕；液体出现浑浊、沉淀时需重新配制。
二、致畸试验胎鼠骨骼完整染色操作流程
1、样本预处理(大鼠、小鼠适用)：
1)取每窝1/2剖宫活胎仔，置于95%乙醇密封固定2-3周；
2)流水冲净固定液后，放入致畸试验骨组织透明液 (KOH溶液) (B1529929) 浸泡8-72h，直至软组织半透明、骨骼轮廓清晰。
注：家兔致畸试验不可分半取样，全部胎仔均需进行全套骨骼染色。
2、骨骼染色：
1)将透明处理的胎仔完全浸没于茜素红应用液中，容器密封避光、室温静置染色6-48h；
2)每日轻摇容器1-2次，直至头骨、四肢骨均匀呈紫红色即可，无需过度延长染色时间。
3、透明脱色(全程避光)：
1)染色结束后取出胎仔，浸入致畸试验骨组织透明液A (KOH-甘油溶液) (B1529923) 避光浸泡1-2d；
2)转入致畸试验骨组织透明液B (甘油溶液) (B1529922) 避光浸泡2-3d；
3)若软组织紫红色残留较重，可适度延长透明处理时长。
4、镜检观察与统计规范
1)标本脱色完成后转移至纯甘油中长期封存，方便镜检与标本留存；
2)采用透射光源，体视显微镜下依次检查颅骨、脊柱、肋骨、胸骨、四肢骨，记录骨化缺失、骨骼融合、肋骨分叉、骨缺损、骨化发育不全等各类骨骼畸形；
3)毒理试验统计要求：所有骨骼畸形、发育异常指标均以窝为单位进行统计，计算总畸胎率、单项骨骼畸形率。
注意事项：
1、稀释必须使用氢氧化钾水溶液，不可用纯水、盐酸、强碱原液直接混合，避免失效、析出沉淀。
2、染色时间需根据标本大小和发育阶段灵活调整，避免染色过深或过浅影响观察。
3、本试剂仅用于整体骨骼钙化染色，不适用于石蜡切片、细胞钙化染色。
4、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Bone Tissue Staining Solution (Alizarin Red Stock Solution) for Teratogenicity Testing is formulated with Alizarin Red, glacial acetic acid, glycerol, chloral hydrate, and stabilizers according to the proportions specified in the national standard. It appears as a clear brown liquid and is slightly irritating. It is suitable for toxicological safety evaluation of food, cosmetics, health products, and industrial chemicals, and for morphological examination of fetal skeletal development in teratogenicity tests involving rats, mice, and rabbits.
Staining Principle: Alizarin Red specifically chelates with calcium salts in bone tissue to form a stable purplish-red complex, completely labeling ossification centers, sutures, ribs, vertebral column, and limb bones throughout the fetal skeleton. After dilution with KOH aqueous solution, it is suitable for whole-mount clearing and staining procedures for fetuses in teratogenicity tests, allowing clear identification of teratogenicity-related skeletal developmental abnormalities such as bone defects, skeletal fusion, bifurcated ribs, and incomplete ossification.
This reagent is intended for research use only and is not suitable for clinical diagnosis or other purposes.
Operating Procedure (For Reference Only):
I. Preparation of Working Solution (Alizarin Red Working Solution)
Take 10 mL of Bone Tissue Staining Solution (Alizarin Red Stock Solution) for Teratogenicity Testing (B1529932), add 10-20 g/L potassium hydroxide (KOH) aqueous solution, and bring the final volume to 1000 mL. Mix thoroughly.
Note: Preparation should be performed in a well-ventilated area while wearing protective gloves and safety goggles. The working solution should be freshly prepared, stored in a brown bottle protected from light, and used within 3 days. If turbidity or precipitation occurs, prepare a fresh working solution.
II. Complete Skeletal Staining Procedure for Fetuses in Teratogenicity Testing
1. Sample Pretreatment (Applicable to Rats and Mice):
1) Select 1/2 of the live fetuses obtained by cesarean section from each litter and fix them in 95% ethanol in a sealed container for 2-3 weeks;
2) After rinsing off the fixative thoroughly under running water, immerse the fetuses in Bone Tissue Clearing Solution (KOH Solution) for Teratogenicity Testing (B1529929) for 8-72 h until the soft tissues become translucent and the skeletal contours are clearly visible.
Note: For rabbit teratogenicity tests, half-litter sampling is not applicable. All fetuses must undergo the complete skeletal staining procedure.
2. Skeletal Staining:
1) Completely immerse the cleared fetuses in the Alizarin Red working solution, seal the container, protect from light, and stain at room temperature for 6-48 h;
2) Gently shake the container 1-2 times daily until the skull and limb bones are uniformly stained purplish-red. Excessive extension of the staining time is unnecessary.
3. Clearing and Decolorization (Protected from Light Throughout):
1) After staining, remove the fetuses and immerse them in Bone Tissue Clearing Solution A (KOH-Glycerol Solution) for Teratogenicity Testing (B1529923), protected from light, for 1-2 d;
2) Transfer the fetuses to Bone Tissue Clearing Solution B (Glycerol Solution) for Teratogenicity Testing (B1529922), protected from light, for 2-3 d;
3) If substantial purplish-red coloration remains in the soft tissues, appropriately extend the clearing time.
4. Microscopic Examination and Statistical Analysis
1) After decolorization is complete, transfer the specimens to pure glycerol for long-term preservation to facilitate microscopic examination and specimen storage;
2) Using transmitted light, sequentially examine the skull, vertebral column, ribs, sternum, and limb bones under a stereomicroscope, and record skeletal abnormalities including absent ossification, skeletal fusion, bifurcated ribs, bone defects, and incomplete ossification;
3) Statistical requirements for toxicological testing: All skeletal malformations and developmental abnormalities should be analyzed on a litter basis, and the total fetal malformation rate and individual skeletal malformation rates should be calculated.
Precautions:
1. Dilution must be performed using potassium hydroxide aqueous solution. Do not directly mix the product with pure water, hydrochloric acid, or concentrated alkaline solution, as this may cause loss of performance or precipitation.
2. The staining time should be adjusted according to specimen size and developmental stage to avoid excessively strong or weak staining that may affect observation.
3. This reagent is intended only for whole-mount skeletal calcification staining and is not suitable for paraffin sections or cellular calcification staining.
4. For your safety and health, wear a laboratory coat and disposable gloves during operation.
规格
规格或纯度
组织学级, 生物染色剂
稳定性与储存
Store at room temperature long term (12 months).
英文名称
Bone Tissue Staining Solution (Alizarin Red Stock Solution) for Teratogenicity Testing
储存条件
室温
运输条件
常规运输
产品线
生物产品
名称和识别符
分子类型
生物试剂/缓冲液
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