优化基于甲酸-福尔马林的大鼠钙骨组织学脱钙方案

S. Amitha Banu, Khan Sharun, Merlin Mamachan, Athira Subash, Vadapalli Deekshita, Kirtika Sharma, Karikalan Mathesh, Obli Rajendran Vinodh kumar, Swapan Kumar Maiti, Abhijit M. Pawde, Laith Abualigah, Kuldeep Dhama, Amarpal
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引用次数: 0

摘要

脱钙在组织学处理中至关重要,尤其是在研究骨骼等矿化组织时。脱钙方法的选择会极大地影响组织切片的质量和组织形态的保存。本研究旨在使用基于甲酸-福尔马林的脱钙溶液建立大鼠腓骨脱钙的标准化方案。根据脱钙时间、甲酸浓度和组织完整性保存等各种参数对该方案进行了系统优化和评估。脱钙过程通过综合评估进行评价,包括大体物理检查、化学分析和放射成像技术。结果表明,浓度为 10%的甲酸对大鼠腓骨样本在 8 天内的脱钙效果最好,在去除矿物质含量的同时还能保持样本结构的完整性。相比之下,浓度为 5%的甲酸无法在十天内完成脱钙,而浓度为 15%的甲酸则会在八天内影响样本质量。组织学分析证实,10% 甲酸浓度在保持组织完整性和达到最佳染色质量方面效果显著。本研究提出的标准化方案为获得一致且高质量的大鼠腓骨组织切片提供了一种有效且可靠的方法。理想的脱钙剂应能有效去除钙盐、保持结构完整性和分子成分、促进快速脱钙但损伤最小,并确保实验室人员易于操作。建议进一步探索其对不同骨类型或物种的适用性,以扩大其研究用途。
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Optimization of Formic Acid-Formalin-Based Decalcification Protocol for Rat Calvarial Bone Histology
Decalcification is crucial in histological processing, particularly for studying mineralized tissues like bone. The choice of decalcification method can significantly impact the quality of histological sections and the preservation of tissue morphology. This study aims to establish a standardized protocol for decalcifying rat calvarial bone using a formic acid-formalin-based decalcification solution. The protocol was systematically optimized and evaluated based on various parameters, including decalcification time, formic acid concentration, and tissue integrity preservation. The decalcification process was evaluated through comprehensive assessments, including gross physical examination, chemical analysis, and radiographic imaging techniques. Our result demonstrated that the 10% formic acid concentration proved most effective for decalcifying rat calvarial bone samples within eight days, excelling in mineral content removal while preserving specimen structural integrity. In contrast, the 5% concentration failed to complete decalcification within ten days, and the 15% compromised sample quality within eight days. Histological analyses confirmed the efficacy of the 10% formic acid concentration in maintaining tissue integrity and achieving optimal staining quality. The standardized protocol presented in this study provides an effective and reliable approach for achieving consistent and high-quality histological sections of rat calvarial bone. An ideal decalcification agent should effectively remove calcium salts, preserve structural integrity and molecular components, facilitate rapid yet minimally damaging decalcification, and ensure ease of handling for laboratory personnel. Further exploration of its applicability to different bone types or species is recommended to broaden its research utility.
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来源期刊
Journal of Experimental Biology and Agricultural Sciences
Journal of Experimental Biology and Agricultural Sciences Agricultural and Biological Sciences-Agricultural and Biological Sciences (all)
CiteScore
1.00
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0.00%
发文量
127
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