BioReagent BioReagent for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
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During routine tissue sectioning, tissues containing bone or calcified lesions cannot be directly embedded in paraffin for sectioning. Due to the density difference between calcium and paraffin, intact sections are difficult to obtain. For calcium-containing tissues, decalcification should be performed after tissue fixation, or fixation and decalcification can be carried out simultaneously, followed by subsequent procedures including dehydration, clearing, infiltration, embedding and sectioning.A variety of decalcifying reagents are available, including organic acids, inorganic acids, ethylenediaminetetraacetic acid (EDTA) and electrolytic decalcification. As a superior chelating decalcifier, EDTA causes minimal damage to tissue structure and well preserves endogenous enzymes within tissues. Tissues decalcified with EDTA are compatible with immunohistochemistry and in situ hybridization staining. However, EDTA decalcification is extremely time-consuming, typically requiring weeks to several months to complete.
JYBL-Ⅰ Decalcifying Solution is mainly composed of dilute acid, formaldehyde, formic acid and other components. Its advantages are as follows:
① Fast decalcification, which can be finished within 24 to 36 hours;
② Slight damage to tissue structure;
③ Thorough decalcification.This product is a rapid decalcifying reagent recommended for decalcification of routine pathological bone tissue samples with low tissue damage. Chemical methods are not suitable for determining the decalcification endpoint for this product.Storage Conditions
Storage Conditions:
Store at room temperature; valid for 1 year.
Required Self-prepared Materials:
1. PBS
2. Distilled water
3. Heating device or microwave oven
Operating Procedures (For Reference Only):
1. Avoid overly thick sampling when preparing bone tissue for decalcification.
2. Rinse fixed tissues with PBS.
3. Wash tissues with distilled water.
4. Immerse tissues in JYBL-Ⅰ Decalcifying Solution for decalcification. Incubate at 37 ℃ to accelerate the decalcification process if needed.
5. Rinse the tissues several times with distilled water.
6. Perform standard dehydration and embedding procedures.
Precautions:
1. Decalcification generally takes 24-36 hours for bone tissue blocks with a thickness of 5 mm.
2. Moderate heating accelerates decalcification; the temperature should not exceed 37-40 ℃. Excessively high temperature will lead to loose and decomposed bone tissue, and the temperature must never go above 60 ℃.
3.Ensure complete decalcification to avoid insufficient or over-decalcification. Minimize decalcification duration while guaranteeing qualified sectioning quality to prevent tissue damage caused by prolonged decalcification.
4.Do not use metal containers during decalcification; glass containers are highly recommended.
5.Bone tissue must be fixed prior to decalcification, or fixation and decalcification shall be conducted concurrently. 5.Decalcification before fixation is forbidden to reduce tissue damage.
6.Check the decalcification progress regularly. Over-decalcification will aggravate tissue damage and interfere with staining outcomes.
7.Wear a lab coat and disposable gloves during operation for personal safety and health protection.
Determination of Decalcification Endpoint (Physical Method):
Use acupuncture, pinching or clamping to test tissues. Terminate decalcification when the bone tissue softens or no resistance is felt during needling. This physical detection method may damage tissue structure, so excessive force should be avoided.
Comprehensive hazard, handling, storage, and regulatory compliance document.
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