Journal Article

Nanoscale imaging of clinical specimens using pathology-optimized expansion microscopy

Chen, Fei, Glass, Benjamin, Boyden, Edward S, Zhao, Yongxin, Irshad, Humayun, Torous, Vanda, Stancu, Andreea L, Stillman, Isaac E, Bucur, Octavian, Weins, Astrid, Oh, Eun-Young, Schnitt, Stuart J, DiStasio, Marcello, Beck, Andrew H

2017

Overview

Expansion microscopy, a technique for super-resolution imaging, is extended to clinical human tissue samples that are formalin fixed, paraffin embedded, stained and/or fresh frozen. 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.

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: 13/51 / 14/1 / 14/105 / 14/32 / 14/56 / 14/63 / 631/1647/328/2238 / 692/53/2421 / 692/700/139/422 / Agriculture / Bioinformatics / Biomedical Engineering/Biotechnology / Biomedicine / Biomolecules / Biopsy / Biotechnology / Breast - diagnostic imaging / Breast - pathology / Breast - ultrastructure / Breast cancer / Computer applications / Deoxyribonucleic acid / Diagnostic imaging / Diffraction / DNA / Electron microscopy / Female / Fidelity / Fluorescence / Formaldehyde / Histological Techniques / Human tissues / Humans / Image Processing, Computer-Assisted - methods / Kidney - diagnostic imaging / Kidney - pathology / Kidney - ultrastructure / Kidneys / Life Sciences / Light microscopy / Medical imaging / Methods / Microscopes / Microscopy / Microscopy - methods / Molecular Imaging - methods / Nanomaterials / Nanomedicine - methods / Nephrosis, Lipoid - diagnostic imaging / Nephrosis, Lipoid - pathology / Optical microscopy / Paraffin / Pathology / Reagents / Samples / Tissues