Wang Shuoshuo
Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
Cancer Program, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
J Histotechnol. 2025 Mar;48(1):68-77. doi: 10.1080/01478885.2024.2446038. Epub 2024 Dec 26.
Bone tissue poses critical roadblocks for spatial transcriptomics and molecular pathology due to a combination of its dense, calcified matrix and inadequate preservation of biomolecules in conventional decalcification. Decalcification is a complex and nuanced histological process to concomitantly preserve nucleic acids, proteins, and tissue architecture, ensuring molecular integrity for downstream assays. However, commonly used agents like formic and hydrochloric acids, while efficient, can degrade biomolecules to varying extents, complicating assays such as PCR, sequencing, immunohistochemistry, and hybridization. Advances in spatial transcriptomics, both sequencing- and imaging-based, emphasize the importance of optimizing decalcification protocols to improve research outcomes. This synoptic and perspective article explores traditional and modern decalcification methods, offering recommendations on technical and methodological refinements for achieving molecularly robust processing of bone and calcified tissues in spatial transcriptomics and molecular pathology.
由于骨组织致密的钙化基质以及传统脱钙过程中生物分子保存不充分，它给空间转录组学和分子病理学带来了严峻的障碍。脱钙是一个复杂且微妙的组织学过程，要同时保存核酸、蛋白质和组织结构，确保下游检测的分子完整性。然而，常用的试剂如甲酸和盐酸虽然有效，但会在不同程度上降解生物分子，使诸如聚合酶链反应（PCR）、测序、免疫组织化学和杂交等检测变得复杂。基于测序和成像的空间转录组学的进展强调了优化脱钙方案以改善研究结果的重要性。这篇综述性和前瞻性文章探讨了传统和现代脱钙方法，为在空间转录组学和分子病理学中实现对骨组织和钙化组织进行分子稳健处理的技术和方法改进提供建议。
