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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.
Comprehensive hazard, handling, storage, and regulatory compliance document.
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