Ugwoke Chiedozie Kenneth, Radochová Barbora, Janáček Jiří, Serša Igor, Ganc Patrik, Cvetko Erika, Umek Nejc
Institute of Anatomy, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia.
Laboratory of Biomathematics, Institute of Physiology of the Czech Academy of Sciences, Prague, Czech Republic.
Microsc Res Tech. 2025 Nov;88(11):3037-3049. doi: 10.1002/jemt.70035. Epub 2025 Jul 7.
Traditional histological methods provide limited insights into the complex 3D microstructure of fascia and its relationship to disease. This study explored the capacity of different 3D microscopy techniques for characterizing the microstructure of fascia lata (FL) and thoracolumbar fascia (TLF). Tissues from four donors were studied using optical projection tomography (OPT), digital light sheet (DLS) microscopy, and magnetic resonance microscopy (MRM). Samples for OPT and DLS were imaged with a custom OPT scanner and the DLS arm of a Leica Stellaris microscope, respectively. MRM was performed using a 9.4 T superconducting magnet and an NMR/MRI spectrometer. Reference histological evaluation was performed to guide the interpretation of 3D data. Image analyses were performed using FIJI and Ellipse software. DLS offered superior resolution, but all techniques revealed a trilaminar structure in both fasciae: a thick, collagen-rich intermediate layer flanked by thinner layers with loose connective tissue. The FL intermediate layer was thinner (210.5-258.7 μm) with longitudinally oriented collagen, while the TLF intermediate layer was thicker (302.3-343.6 μm) with both oblique and longitudinal fibers. The superficial layer in FL was thicker (128.8-161.5 μm) than in TLF (84.65-123.10 μm) across imaging modalities. The deep layer also varied between fasciae, with 54.3-73.8 μm in FL and 44.78-70.30 μm in TLF. Layer thickness measurements did not differ significantly across techniques. This study demonstrates the feasibility of different 3D microscopy techniques for visualizing and quantifying fascia extracellular matrix structure and organization, laying the groundwork for future investigations into potential structural alterations in disease.
传统组织学方法对筋膜复杂的三维微观结构及其与疾病的关系提供的见解有限。本研究探索了不同三维显微镜技术表征阔筋膜（FL）和胸腰筋膜（TLF）微观结构的能力。使用光学投影断层扫描（OPT）、数字光片（DLS）显微镜和磁共振显微镜（MRM）对来自四名供体的组织进行了研究。OPT和DLS的样本分别使用定制的OPT扫描仪和徕卡Stellaris显微镜的DLS臂进行成像。MRM使用9.4 T超导磁体和NMR/MRI光谱仪进行。进行了参考组织学评估以指导对三维数据的解释。使用FIJI和Ellipse软件进行图像分析。DLS提供了更高的分辨率，但所有技术都揭示了两种筋膜的三层结构：中间是一层厚厚的、富含胶原蛋白的层，两侧是较薄的、含有疏松结缔组织的层。FL中间层较薄（210.5 - 258.7μm），胶原蛋白呈纵向排列，而TLF中间层较厚（302.3 - 343.6μm），含有斜向和纵向纤维。在所有成像方式中，FL的表层（128.8 - 161.5μm）比TLF的表层（84.65 - 123.10μm）更厚。深层在两种筋膜之间也有所不同，FL为54.3 - 73.8μm，TLF为44.78 - 70.30μm。各技术之间的层厚度测量结果无显著差异。本研究证明了不同三维显微镜技术用于可视化和量化筋膜细胞外基质结构与组织的可行性，为未来对疾病潜在结构改变的研究奠定了基础。
