Expansion-enhanced super-resolution radial fluctuations enable nanoscale molecular profiling of pathology specimens

Expansion microscopy physically enlarges biological specimens to achieve nanoscale resolution using diffraction-limited microscopy systems 1 . However, optimal performance is usually reached using laser-based systems (for example, confocal microscopy), restricting its broad applicability in clinical...

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Bibliographic Details
Published in:	Nature nanotechnology Vol. 18; no. 4; pp. 336 - 342
Main Authors:	Kylies, Dominik, Zimmermann, Marina, Haas, Fabian, Schwerk, Maria, Kuehl, Malte, Brehler, Michael, Czogalla, Jan, Hernandez, Lola C., Konczalla, Leonie, Okabayashi, Yusuke, Menzel, Julia, Edenhofer, Ilka, Mezher, Sam, Aypek, Hande, Dumoulin, Bernhard, Wu, Hui, Hofmann, Smilla, Kretz, Oliver, Wanner, Nicola, Tomas, Nicola M., Krasemann, Susanne, Glatzel, Markus, Kuppe, Christoph, Kramann, Rafael, Banjanin, Bella, Schneider, Rebekka K., Urbschat, Christopher, Arck, Petra, Gagliani, Nicola, van Zandvoort, Marc, Wiech, Thorsten, Grahammer, Florian, Sáez, Pablo J., Wong, Milagros N., Bonn, Stefan, Huber, Tobias B., Puelles, Victor G.
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 01-04-2023 Nature Publishing Group
Subjects:	631/61/350/2093 639/925/930/328/2238 Animals Biopsy Chemistry and Materials Science Computer applications Confocal microscopy Endoplasmic reticulum Fluctuations Humans Image Enhancement Image processing Image resolution Ischemia Kidney Kidneys Laser applications Letter Light emitting diodes Materials Science Mice Microscopy Microscopy, Confocal - methods Microscopy, Fluorescence - methods Nanotechnology Nanotechnology and Microengineering Paraffin Pathology Workflow
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Summary:	Expansion microscopy physically enlarges biological specimens to achieve nanoscale resolution using diffraction-limited microscopy systems 1 . However, optimal performance is usually reached using laser-based systems (for example, confocal microscopy), restricting its broad applicability in clinical pathology, as most centres have access only to light-emitting diode (LED)-based widefield systems. As a possible alternative, a computational method for image resolution enhancement, namely, super-resolution radial fluctuations (SRRF) 2 , 3 , has recently been developed. However, this method has not been explored in pathology specimens to date, because on its own, it does not achieve sufficient resolution for routine clinical use. Here, we report expansion-enhanced super-resolution radial fluctuations (ExSRRF), a simple, robust, scalable and accessible workflow that provides a resolution of up to 25 nm using LED-based widefield microscopy. ExSRRF enables molecular profiling of subcellular structures from archival formalin-fixed paraffin-embedded tissues in complex clinical and experimental specimens, including ischaemic, degenerative, neoplastic, genetic and immune-mediated disorders. Furthermore, as examples of its potential application to experimental and clinical pathology, we show that ExSRRF can be used to identify and quantify classical features of endoplasmic reticulum stress in the murine ischaemic kidney and diagnostic ultrastructural features in human kidney biopsies. Complex clinical samples can be imaged with 25 nm resolution using a light-emitting-diode-based wide-field microscope.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1748-3387 1748-3395
DOI:	10.1038/s41565-023-01328-z