Expansion microscopy (ExM), a method for improving the resolution of light microscopy by physically expanding a specimen, has not been applied to clinical tissue samples. Here we report a clinically optimized form of ExM that supports nanoscale imaging of human tissue specimens that have been fixed with formalin, embedded in paraffin, stained with hematoxylin and eosin, and/or fresh frozen. The method, which we call expansion pathology (ExPath), converts clinical samples into an ExM-compatible state, then applies an ExM protocol with protein anchoring and mechanical homogenization steps optimized for clinical samples. ExPath enables ~70-nm-resolution imaging of diverse biomolecules in intact tissues using conventional diffraction-limited microscopes and standard antibody and fluorescent DNA in situ hybridization reagents. We use ExPath for optical diagnosis of kidney minimal-change disease, a process that previously required electron microscopy, and we demonstrate high-fidelity computational discrimination between early breast neoplastic lesions for which pathologists often disagree in classification. ExPath may enable the routine use of nanoscale imaging in pathology and clinical research.

中文翻译：

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使用病理优化的扩展显微镜对临床标本进行纳米成像

扩展显微镜（ExM）是一种通过物理扩展标本来提高光学显微镜分辨率的方法，尚未应用于临床组织样本。在这里，我们报告了ExM的临床优化形式，该形式支持已用福尔马林固定，石蜡包埋，苏木精和曙红染色和/或新鲜冷冻的人体组织标本的纳米级成像。我们将这种方法称为扩展病理学（ExPath），将临床样本转换为与ExM兼容的状态，然后应用具有针对临床样本优化的蛋白质锚固和机械均质化步骤的ExM协议。ExPath可使用常规的衍射极限显微镜以及标准抗体和原位荧光DNA ，在完整组织中对各种生物分子进行约70 nm分辨率的成像杂交试剂。我们将ExPath用于肾脏最小变化疾病的光学诊断，这是以前需要电子显微镜检查的过程，并且我们证明了早期乳腺肿瘤性病变之间的高保真度计算区别，病理学家通常对此并不认同。ExPath可以在病理学和临床研究中常规使用纳米级成像。