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。EDTA是一种相对较好的螯合脱钙剂，对组织结构影响最小，可以较好的保存组织的某些酶类，经EDTA脱钙后的组织可以进行免疫组化和原位杂交染色，但是该法脱钙速度太慢，一般脱需要数周至数月。
JYBL-Ⅲ脱钙液主要由甲醛、50%甲酸等组成，其优点是：
①甲酸是最接近无机酸和有机酸临界线的一种有机酸，对组织的结构损害小；
②脱钙完全。
该脱钙液适用于病理科特殊骨组织标本脱钙，如果在脱钙前对组织固定，效果更好。
该试剂仅用于科研领域，不适用于临床诊断或其他用途。
系列产品概览
| 货号 | 名称 | 主要成分 | 脱钙时间 | 组织损伤风险 | 核心优势 | 适用标本类型 |
| J1525466 | JYBL-Ⅰ脱钙液 | 稀酸、甲醛、甲酸 | 24～36 h | 相对较小 | 速度与损伤的基础平衡 | 常规骨组织标本 |
| J1529419 | JYBL-Ⅱ脱钙液 | 稀酸、甲醛、甲酸 | 约 24 h（比Ⅰ更快） | 有一定损伤（比Ⅰ略高） | 速度优于Ⅰ | 常规骨组织标本 |
| J1529418 | JYBL-Ⅲ脱钙液 | 甲醛、50%甲酸 | 室温：2～3 天 | 相对最小（甲酸最温和） | 甲酸对组织损伤小，形态保存更佳 | 特殊骨组织标本 |
| J1529417 | JYBL-Ⅳ脱钙液 | 甲醛、甲酸（不含强酸） | 37℃加热：2～3 h
室温：2～3 天 | 相对较小，加热需控温 | 速度最快，专为骨穿刺微量样本设计 | 骨组织穿刺样本 |
自备材料：
1、蒸馏水或PBS
2、加热装置或微波炉、脱钙专用容器（玻璃）
操作步骤（仅供参考）：
1、骨组织脱钙时，取材不宜过厚，一般大约 3～5 mm。
2、组织固定后，用蒸馏水或PBS清洗3次，每次 20 min。
3、组织转移至 20～30 倍体积的JYBL-Ⅲ脱钙液中，脱钙 2～3 天；如果想加快脱钙速度，可以置于 37℃ 进行脱钙。
4、用蒸馏水冲洗数次。
5、常规脱水、包埋。
注意事项：
1、厚度 5 mm 的骨组织块脱钙时间一般脱钙 2-3 天即可。
2、适当加温能加快脱钙的速度，一般不应超过 37～40℃，温度过高容易使骨组织造成松散解体，尤其不可大于 60℃。
3、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。
4、脱钙用具避免使用金属容器，尽量使用玻璃容器。
5、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。
6、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。
7、为了您的安全和健康，请穿实验服并戴一次性手套操作。
附录：
脱钙终点的测定（物理法）：采用针刺、手掐、钳夹等方法，当骨组织变软或针刺时没有阻力感即可终止脱钙。物理检测法会对组织结构有一定的损害，尽量避免用力过大或反复检测。
During tissue sectioning, some tissues contain bone or calcified foci. Calcified tissues are not suitable for direct paraffin embedding and sectioning, because the density difference between calcium and paraffin makes it difficult to obtain intact sections. For calcified tissues, it is best to fix first and then decalcify, or perform both simultaneously, followed by downstream procedures such as dehydration, clearing, paraffin infiltration, embedding, and sectioning. Many reagents are available for decalcification, including organic acids, inorganic acids, ethylenediaminetetraacetic acid (EDTA), and electrolytic decalcification. EDTA is a relatively good chelating decalcifying agent, with minimal impact on tissue structure and good preservation of certain enzymes in tissues. Tissues decalcified with EDTA can be used for immunohistochemistry and in situ hybridization staining. However, this method is too slow, generally taking several weeks to months.
JYBL-Ⅲ Decalcifying Solution is mainly composed of formaldehyde, 50% formic acid, etc. Its advantages are:
① formic acid is an organic acid that is closest to the critical line between inorganic and organic acids, causing minimal damage to tissue structure;
② complete decalcification.
This decalcifying solution is used for decalcification of special bone tissue specimens in pathology departments. If the tissue is fixed before decalcification, the effect is better.
This reagent is intended for research use only and is not suitable for clinical diagnosis or other applications.
Product Series Overview
| Cat. No. | Product Name | Main Components | Decalcification Time | Tissue Damage Risk | Core Advantages | Applicable Specimen Types |
| J1525466 | JYBL-Ⅰ Decalcifying Solution | Dilute acid, formaldehyde, formic acid | 24–36 h | Relatively low | Balanced speed and tissue preservation | Routine bone tissue specimens |
| J1529419 | JYBL-Ⅱ Decalcifying Solution | Dilute acid, formaldehyde, formic acid | Approx. 24 h (faster than Ⅰ) | Moderate (slightly higher than Ⅰ) | Faster decalcification than Ⅰ | Routine bone tissue specimens |
| J1529418 | JYBL-Ⅲ Decalcifying Solution | Formaldehyde, 50% formic acid | Room temperature: 2–3 days | Relatively minimal (formic acid is the mildest) | Minimal tissue damage; superior morphology preservation | Special bone tissue specimens |
| J1529417 | JYBL-Ⅳ Decalcifying Solution | Formaldehyde, formic acid (acid-free formulation) | 37°C incubation: 2–3 h | Relatively low; temperature control required | Fastest decalcification; designed for bone marrow biopsy | Bone marrow biopsy specimens |
Materials Required (Self-Prepared):
1.Distilled water or PBS
2.Heating apparatus or microwave oven, and dedicated decalcification container (glass)
Operating Instructions (For Reference Only):
1.For bone tissue decalcification, the specimen should not be too thick; generally approximately 3–5 mm.
2.After fixation, wash the tissue 3 times with distilled water or PBS, 20 minutes each time.
3.Transfer the tissue to 20–30 volumes of JYBL-Ⅲ Decalcifying Solution, and decalcify for 2–3 days; to accelerate decalcification, it can be performed at 37°C.
4.Rinse several times with distilled water.
5.Proceed with routine dehydration and embedding.
Precautions:
1.For bone tissue blocks of 5 mm thickness, decalcification generally takes 2–3 days.
2.Moderate heating can accelerate decalcification, but the temperature should generally not exceed 37–40°C. Excessive temperatures may cause loosening and disintegration of bone tissue; temperatures above 60°C must not be used.
3.Decalcification should be thorough to prevent under-decalcification or over-decalcification. The degree of decalcification should be controlled to ensure that tissue sectioning is not affected while shortening the decalcification time as much as possible, to avoid tissue damage caused by prolonged decalcification.
4.Avoid using metal containers for decalcification; glass containers are recommended.
5.Bone tissue should be fixed before decalcification, or fixation and decalcification performed simultaneously. Decalcification should not be performed before fixation, in order to reduce tissue damage.
6.Check the degree of decalcification periodically. Over-decalcification will increase tissue damage and affect staining results.
7.For your safety and health, please wear a lab coat and disposable gloves during handling.
Appendix:
Determination of Decalcification Endpoint (Physical Method): Use methods such as needling, pinching, or clamping. Decalcification can be terminated when the bone tissue becomes soft or when no resistance is felt upon needling. Physical detection methods may cause certain damage to tissue structure; therefore, excessive force or repeated testing should be avoided.
ZJ26F1040100
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