Department of Physics and Munich School of BioEngineering, Technical University of Munich, 85748 Garching, Germany;
Department of Physics and Munich School of BioEngineering, Technical University of Munich, 85748 Garching, Germany.
Proc Natl Acad Sci U S A. 2018 Mar 6;115(10):2293-2298. doi: 10.1073/pnas.1720862115. Epub 2018 Feb 20.
Many histological methods require staining of the cytoplasm, which provides instrumental details for diagnosis. One major limitation is the production of 2D images obtained by destructive preparation of 3D tissue samples. X-ray absorption micro- and nanocomputed tomography (microCT and nanoCT) allows for a nondestructive investigation of a 3D tissue sample, and thus aids to determine regions of interest for further histological examinations. However, application of microCT and nanoCT to biological samples (e.g., biopsies) is limited by the missing contrast within soft tissue, which is important to visualize morphological details. We describe an eosin-based preparation overcoming the challenges of contrast enhancement and selectivity for certain tissues. The eosin-based staining protocol is suitable for whole-organ staining, which then enables high-resolution microCT imaging of whole organs and nanoCT imaging of smaller tissue pieces retrieved from the original sample. Our results demonstrate suitability of the eosin-based staining method for diagnostic screening of 3D tissue samples without impeding further diagnostics through histological methods.
许多组织学方法需要对细胞质进行染色，这为诊断提供了仪器细节。一个主要的限制是通过对 3D 组织样本进行破坏性制备来获得 2D 图像。X 射线吸收微纳计算机断层扫描（microCT 和 nanoCT）允许对 3D 组织样本进行非破坏性研究，从而有助于确定进一步组织学检查的感兴趣区域。然而，微 CT 和纳 CT 在生物样本（例如活检）中的应用受到软组织内缺失对比度的限制，这对于可视化形态细节很重要。我们描述了一种基于曙红的制备方法，该方法克服了对比度增强和对某些组织选择性的挑战。基于曙红的染色方案适用于整个器官的染色，这使得整个器官的高分辨率 microCT 成像和从原始样本中取出的较小组织块的 nanoCT 成像成为可能。我们的结果表明，基于曙红的染色方法适用于 3D 组织样本的诊断筛选，而不会通过组织学方法阻碍进一步的诊断。
