Buxton Andrew W, Jalufka Frank L, Hruska Margaret E, Kubaney Jack R, McCreedy Dylan A
Department of Biomedical Engineering, Texas A&M University, College Station, TX, United States.
Department of Biology, Texas A&M University, College Station, TX, United States.
Front Neurosci. 2025 Jun 13;19:1601360. doi: 10.3389/fnins.2025.1601360. eCollection 2025.
Tissue clearing and 3D imaging have emerged as powerful techniques to assess the cellular and tissue-level architecture of the spinal cord. With the rapidly increasing variety and complexity of optical tissue clearing techniques, there is a critical need for optimization and streamlining of tissue-specific protocols, particularly when dealing with injury or disease states. We evaluated and combined multiple organic solvent-based techniques to develop sciDISCO: a spinal cord injury-optimized DISCO tissue clearing protocol. sciDISCO allows for the robust clearing, labeling, and 3D imaging of the intact spinal cord, as well as clearing around and through the lesion site formed after contusive spinal cord injury. In addition, we have identified alternatives for hazardous chemicals commonly used in organic solvent-based clearing including dichloromethane and dibenzyl ether. In this study, we demonstrate the compatibility of sciDISCO with multiple different labeling techniques to provide robust analysis of unique neuronal populations and morphologies in addition to cellular and tissue-level changes occurring following spinal cord injury.
组织透明化和三维成像已成为评估脊髓细胞和组织水平结构的强大技术。随着光学组织透明化技术的种类和复杂性迅速增加，迫切需要优化和简化针对特定组织的方案，尤其是在处理损伤或疾病状态时。我们评估并结合了多种基于有机溶剂的技术，开发出了sciDISCO：一种针对脊髓损伤优化的DISCO组织透明化方案。sciDISCO能够对完整脊髓进行强有力的透明化、标记和三维成像，还能对脊髓挫伤损伤后形成的损伤部位及其周围进行透明化处理。此外，我们还找到了常用于基于有机溶剂的透明化处理的危险化学品（如二氯甲烷和二苄醚）的替代品。在本研究中，我们证明了sciDISCO与多种不同标记技术的兼容性，除了脊髓损伤后发生的细胞和组织水平变化外，还能对独特的神经元群体和形态进行强有力的分析。
