在组织切片过程中一些组织内含有骨质或钙化灶时，含钙的组织不宜直接用石蜡包埋切片，这是因为钙和石蜡之间的密度不同，较难切出完整的切片。对含钙组织最好固定之后，再进行脱钙或二者同时进行，然后进行下游操作如脱水、透明、浸蜡、包埋、切片。用于脱钙的试剂很多，脱钙剂包括有机酸、无机酸、乙二胺四乙酸 (EDTA) 以及电解法脱钙，在有机酸中，甲酸、乙酸比较适合于骨髓组织脱钙。
电解脱钙液主要由甲酸、稀酸等组成；其优点：①脱钙速度快；②适用于大多数组织的脱钙；其缺点：①对组织有一定的损害；②脱钙后需要硫酸钠溶液进行碱处理。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、PBS
2、蒸馏水
3、恒温水浴箱
4、5% 硫酸钠溶液
操作步骤 (仅供参考)：
1、骨组织脱钙时，取材不易过厚，一般大约 5mm。
2、组织固定后，用 PBS 清洗 3 次，每次 20min。
3、组织用蒸馏水清洗 3 次，每次 20min。
4、安装电解装置：取铂金丝一根，将标本用铂金丝一端缠绕数圈，置于容器底部一侧，铂金丝另一端连接电源正极；在容器的另一侧将一只钢片或炭精棒的下端插入容器内，上段与电源负极相连；在正负极之间用一块绝缘的有机玻璃板隔开以防止电极接触引起导电。
5、加入 150～200ml 电解脱钙液，置于 37～45℃ 恒温水浴箱中，一般采用直流电源 6V，如有整流器可调至 8～10V；一般情况下电解 10min～3h，密质骨可延长至 6～8h。
6、用流水冲洗数小时。
7、入 5% 硫酸钠溶液进行碱处理。
8、常规脱水、包埋。
注意事项：
1、厚度 5mm 的骨组织块脱钙时间：一般脱钙10min～3h 即可。
2、脱钙应彻底，防止脱钙不足或过度，脱钙程度应控制在不影响组织切片的同时尽量缩短脱钙时间，以免脱钙过长引起组织损害。
3、骨组织脱钙应先固定后脱钙或脱钙固定同时进行，不应先脱钙后固定，以便减少组织的损伤程度。
4、每隔一段时间检测一次脱钙程度，脱钙过度会增加组织的损伤程度，影响染色结果。
5、为了您的安全和健康，请穿实验服并戴一次性手套操作。
When certain tissues contain bone substance or calcified lesions during tissue sectioning, calcium-containing tissues cannot be directly embedded in paraffin for sectioning. This is attributed to the density difference between calcium and paraffin, which makes it difficult to obtain intact sections.For calcium-containing tissues, decalcification is preferably performed after fixation, or fixation and decalcification may be carried out simultaneously, followed by subsequent procedures including dehydration, clearing, wax impregnation, embedding and sectioning.
A variety of decalcifying reagents are available, including organic acids, inorganic acids, ethylenediaminetetraacetic acid (EDTA) and electrolytic decalcification methods. Among organic acids, formic acid and acetic acid are suitable for decalcification of bone marrow tissues.
Electrolytic Decalcifying Solution is mainly composed of formic acid and dilute acid.Advantages: ① Rapid decalcification speed; ② Applicable to decalcification of most types of tissues.
Disadvantages: ① It may cause certain damage to tissues; ② Alkali treatment with sodium sulfate solution is required after decalcification.This reagent is intended solely for scientific research and shall not be used for clinical diagnosis or other purposes.
Materials to be Prepared by User：
1. PBS
2. Distilled water
3. Thermostatic water bath
4. 5% Sodium Sulfate Solution
Operating Procedures (For Reference Only)：
1. When decalcifying bone tissues, the sampled tissue should not be too thick, with a recommended thickness of approximately 5 mm.
2. After tissue fixation, rinse the sample with PBS 3 times, 20 minutes each time.
3. Rinse the tissue with distilled water 3 times, 20 minutes each time.
4. Assemble the electrolytic device: Take a platinum wire, wrap one end of the wire around the specimen and place it on one side of the container bottom, then connect the other end of the platinum wire to the positive electrode of the power supply. On the opposite side of the container, insert the lower end of a steel sheet or carbon rod into the solution, and connect its upper part to the negative electrode of the power supply. Place an insulated organic glass plate between the positive and negative electrodes to prevent electrical conduction caused by electrode contact.
5. Add 150～200 mL Electrolytic Decalcification Solution and place the container in a thermostatic water bath at 37～45 ℃. A 6 V direct current power supply is generally used; the voltage can be adjusted to 8～10 V if a rectifier is equipped. Electrolysis normally lasts 10 minutes to 3 hours, and the duration can be extended to 6～8 hours for compact bone.
6. Rinse the sample with running tap water for several hours.
7. Perform alkali treatment by immersing the sample in 5% Sodium Sulfate Solution.
8. Conduct routine dehydration and embedding.
Precautions：
1. For bone tissue blocks with a thickness of 5 mm, the decalcification period is generally 10 min to 3 h.
2. Complete decalcification is required to avoid insufficient or excessive decalcification. The decalcification degree shall be controlled to ensure no adverse impact on tissue sectioning while minimizing decalcification duration, so as to prevent tissue damage caused by overlong decalcification.
3. Bone tissue shall be fixed prior to decalcification, or fixation and decalcification shall be conducted concurrently. Decalcification before fixation is prohibited to reduce tissue damage.
4. Check the decalcification progress at regular intervals. Excessive decalcification will aggravate tissue damage and compromise staining results.
5. Wear a lab coat and disposable gloves during operation for personal safety and health.
ZJ26F0939209
危险品化学品经营许可证（带存储）