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致畸试验骨组织染色液 (茜素红应用液Ⅱ)
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概述
致畸试验骨组织染色液 (茜素红应用液Ⅱ) 为即用型骨骼染色工作液，由茜素红、氢氧化钾为核心组分复配而成，液体呈红棕色，具备轻微腐蚀性。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，专门用于大鼠、小鼠、家兔致畸试验中去皮去内脏并完成透明处理后的胎仔整体骨骼染色。
染色原理：茜素红可特异性与骨骼内钙盐螯合，生成稳定紫红色复合物，完整标记胎仔头骨、肋骨、胸骨、脊柱、四肢骨的骨化结构，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)：
1、样本预处理(大鼠、小鼠适用)：
1)取每窝 1/2 剖宫活胎仔，小心完整剥皮、去除内脏与全身脂肪，重点清理颈肩区域脂肪组织，操作轻柔避免肋骨、四肢骨断裂。
2)将处理后的胎仔放入致畸试验骨组织透明液(KOH溶液)(B1529929)浸泡 8-72h，直至软组织完全透明、骨骼轮廓清晰。
注：家兔致畸试验不可分半取样，全部胎仔均需完整染色检查。
2、骨骼染色(本品直接使用，无需稀释)：
1)将去皮透明完成的胎仔完全浸没于致畸试验骨组织染色液(茜素红应用液Ⅱ)(B1529931)中，密封避光室温染色 6-48h；
2)每日轻摇容器 2-3 次，直至全身骨骼均匀呈紫红色即可，无需过度延长染色时间。
3、分级透明脱色(配套试剂需自备)：
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、本品为预配应用液，已添加氢氧化钾，禁止额外加水、酸碱、强碱原液稀释，否则染料析出、染色失效。
2、染色时长可根据胎鼠日龄、标本大小灵活微调，避免染色过深导致软组织背景发红，干扰畸形判断。
3、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。
4、仅限去皮去内脏的透明胎仔染色，不适用于未剥皮完整胎仔、石蜡病理切片、细胞钙化染色、脱钙组织染色。
5、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 Working Solution II) for Teratogenicity Testing is a ready-to-use skeletal staining working solution formulated with Alizarin Red and potassium hydroxide as the core components. The solution is reddish-brown and exhibits slight corrosiveness. It is suitable for toxicological safety evaluation of food, cosmetics, health products, and industrial chemicals, and is specifically intended for whole-mount skeletal staining of fetuses from rats, mice, and rabbits after skinning, evisceration, and clearing in teratogenicity studies.
Staining Principle: Alizarin Red specifically chelates with calcium salts in skeletal tissue to form a stable purplish-red complex, allowing complete visualization of the ossified structures of the fetal skull, ribs, sternum, vertebral column, and limb bones, and facilitating 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):
1. Sample Pretreatment (Applicable to Rats and Mice):
1) Select 1/2 of the live fetuses obtained by cesarean section from each litter. Carefully remove the skin completely, remove the internal organs and body fat, with particular attention to adipose tissue in the neck and shoulder regions. Handle gently to avoid fractures of the ribs and limb bones.
2) Place the treated fetuses in Bone Tissue Clearing Solution (KOH Solution) for Teratogenicity Testing (B1529929) and immerse for 8-72 h until the soft tissues become completely transparent and the skeletal contours are clearly visible.
Note: For rabbit teratogenicity tests, half-litter sampling is not applicable. All fetuses must undergo complete skeletal staining and examination.
2. Skeletal Staining (Use Directly Without Dilution):
1) Completely immerse the skinned and cleared fetuses in Bone Tissue Staining Solution (Alizarin Red Working Solution II) for Teratogenicity Testing (B1529931), seal the container, protect from light, and stain at room temperature for 6-48 h;
2) Gently shake the container 2-3 times daily until the entire skeleton is uniformly stained purplish-red. Excessive extension of the staining time is unnecessary.
3. Graded Clearing and Decolorization (Supporting Reagents Required):
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, examine the skull, vertebral column, ribs, sternum, and limb bones sequentially under a stereomicroscope, and record skeletal abnormalities including absent ossification, skeletal fusion, bifurcated ribs, bone defects, and incomplete ossification;
3) Toxicological test statistics: 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. This product is a ready-to-use working solution containing potassium hydroxide. Do not further dilute it with water, acids, alkalis, or concentrated alkaline solutions, as this may cause dye precipitation and loss of staining performance.
2. The staining time may be adjusted according to fetal age and specimen size. Avoid overstaining, which may cause excessive red background staining of soft tissues and interfere with the evaluation of skeletal abnormalities.
3. The staining duration should be adjusted according to specimen size and developmental stage to avoid excessively strong or weak staining that may affect observation.
4. This product is intended only for staining cleared fetuses after skinning and evisceration. It is not suitable for intact unskinned fetuses, paraffin-embedded histological sections, cellular calcification staining, or staining of decalcified tissues.
5. 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, BioReagent, 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 Working Solution II) 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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