胼胝质（callose）是一类由葡萄糖残基以不同于纤维素的方式连接形成的非纤维素多糖，参与植物筛管代谢、配子体发育等重要生命活动并发挥关键调控作用，其合成与降解过程直接影响植物正常生长代谢，是植物学研究的重要内容。
植物胼胝质染色液（苯胺蓝可见光法）利用苯胺蓝对胼胝质的特异性染色特性，可将其清晰染为蓝色，便于在可见光显微镜下直接观察与分析。本品仅限科研领域使用，不得用于临床诊断及其他非科研用途。
自备材料
- AAF固定液或Carnoy固定液、50%乙醇、无水乙醇、1×PBS
- 新鲜的植物组织
操作步骤（仅供参考）
1. 新鲜植物组织切成2 mm左右的薄片，植物叶片推荐裁剪成1 cm*2 cm，幼嫩植物叶片可直接浸于AAF固定液或Carnoy固定液固定24小时。
2. 使用无水乙醇浸洗两次，每次1分钟，然后转入至少10倍体积的100%乙醇中保存。
3. 染色前取出组织，浸于50%乙醇中平衡30 min，取出稍沥干。
4. 随后浸于1×PBS中平衡30 min，取出稍沥干。
5. 组织切片滴加或浸于胼胝质染色液中，室温避光染色4~24小时。
6. 用蒸馏水冲洗组织切片，小心的将染色后的组织转移至载玻片上，滴加少量甘油明胶封片剂后封片观察。
染色结果
胼胝质呈蓝色。
注意事项
1. 在条件允许下，推荐将浸于固定液中的植物组织负压真空处理20 min，有助于固定液的渗透。
2. 如暂时不进行实验或需同时处理大量样本，浸于100%乙醇的样本可于2-8℃保存至少1周。
3. 样本染色完成后尽快拍照保存结果。
4. 试剂开封后请尽快使用，以防影响后续实验效果。
5. 为了您的安全和健康，请穿实验服并戴一次性手套操作。
Callose is a type of non-cellulosic polysaccharide composed of glucose residues linked in a manner distinct from cellulose. It plays a crucial regulatory role in key biological processes such as plant sieve tube metabolism and gametophyte development. The processes of its synthesis and degradation directly impact normal plant growth and metabolism, making it a significant subject in botanical research.
Plant Callose Staining Solution (Aniline Blue Visible-Light Method) utilizes the specific staining property of aniline blue for callose, clearly staining it blue. This facilitates direct observation and analysis under a visible light microscope. This product is intended for research use only and must not be used for clinical diagnosis or other non-research purposes.
Materials to Be Prepared by User
- AAF Fixative Solution or Carnoy's Fixative Solution, 50% Ethanol, Absolute Ethanol, 1× PBS
- Fresh plant tissue
Procedure (For Reference Only)
Cut fresh plant tissue into slices approximately 2 mm thick. For plant leaves, it is recommended to cut them into pieces about 1 cm * 2 cm. Tender plant leaves can be directly immersed in AAF Fixative Solution or Carnoy's Fixative Solution for 24 hours.
Rinse the tissue twice with absolute ethanol, 1 minute each time. Then transfer and store the tissue in at least 10 times its volume of 100% ethanol.
Before staining, retrieve the tissue and immerse it in 50% ethanol for equilibration for 30 minutes. Remove and allow to drain slightly.
Subsequently, immerse the tissue in 1× PBS for equilibration for 30 minutes. Remove and allow to drain slightly.
Apply the Callose Staining Solution dropwise onto the tissue section or immerse the section in the staining solution. Stain at room temperature, protected from light, for 4~24 hours.
Rinse the tissue section with distilled water. Carefully transfer the stained tissue onto a glass slide. Add a small amount of glycerol-gelatin mounting medium before coverslipping and observation.
Staining Results
Callose appears blue.
Notes
When conditions permit, it is recommended to subject the plant tissue immersed in fixative to negative pressure/vacuum treatment for 20 minutes to aid fixative penetration.
If the experiment is not to be performed immediately or a large number of samples need to be processed simultaneously, samples stored in 100% ethanol can be kept at 2-8°C for at least one week.
Photograph the stained samples as soon as possible after staining to preserve the results.
Use the reagents as soon as possible after opening to avoid affecting subsequent experimental results.
For your safety and health, please wear a lab coat and disposable gloves during operation.
ZJ26F0434138
危险品化学品经营许可证（带存储）