Three-dimensional virtual histology enabled through cytoplasm-specific X-ray stain for microscopic and nanoscopic computed tomography
Three-dimensional virtual histology enabled through cytoplasm-specific X-ray stain for microscopic and nanoscopic computed tomography
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DOI：
10.1073/pnas.1720862115
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发表时间：
2018-03-06
影响因子：
11.1
通讯作者：
Pfeiffer, Franz
中科院分区：
文献类型：
--
作者：
Busse, Madleen;Mueller, Mark;Pfeiffer, Franz
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。一项主要限制是通过破坏性制备 3D 组织样本获得的 2D 图像。 X 射线吸收微米和纳米计算机断层扫描（microCT 和 nanoCT）可对 3D 组织样本进行无损研究，从而有助于确定感兴趣区域以进行进一步的组织学检查。然而，microCT 和 nanoCT 在生物样本（例如活检）中的应用受到软组织内对比度缺失的限制，而软组织内对比度对于可视化形态细节非常重要。我们描述了一种基于伊红的制剂，克服了某些组织的对比度增强和选择性的挑战。基于曙红的染色方案适用于整个器官染色，从而能够对整个器官进行高分辨率 microCT 成像，并对从原始样本中检索的较小组织碎片进行 nanoCT 成像。我们的结果证明基于曙红的染色方法适用于 3D 组织样本的诊断筛查，且不会妨碍通过组织学方法进行进一步的诊断。
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.