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

Dominik Kylies, Marina Zimmermann, Fabian Haas, Maria Schwerk, Malte Kuehl, Michael Brehler, Jan Czogalla, Lola C. Hernandez, Leonie Konczalla, Yusuke Okabayashi, Julia Menzel, Ilka Edenhofer, Sam Mezher, Hande Aypek, Bernhard Dumoulin, Hui Wu, Smilla Hofmann, Oliver Kretz, Nicola Wanner, Nicola M. TomasSusanne Krasemann, Markus Glatzel, Christoph Kuppe, Rafael Kramann, Bella Banjanin, Rebekka K. Schneider, Christopher Urbschat, Petra Arck, Nicola Gagliani, Marc van Zandvoort, Thorsten Wiech, Florian Grahammer, Pablo J. Sáez, Milagros N. Wong, Stefan Bonn, Tobias B. Huber, Victor G. Puelles*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

8 Citations (Scopus)
26 Downloads (Pure)


Expansion microscopy physically enlarges biological specimens to achieve nanoscale resolution using diffraction-limited microscopy systems1. 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.

Original languageEnglish
Pages (from-to)336-342
Number of pages7
JournalNature Nanotechnology
Issue number4
Publication statusPublished - 10 Apr 2023

Bibliographical note

Open access funding provided by Universitätsklinikum
Hamburg-Eppendorf (UKE).

Publisher Copyright:
© 2023, The Author(s).


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

    Kylies, D. (Creator), Zimmermann, M. (Creator), Haas, F. (Creator), Schwerk, M. (Creator), Kuehl, M. (Creator), Brehler, M. (Creator), Czogalla, J. (Creator), Hernandez, L. C. (Creator), Konczalla, L. (Creator), Okabayashi, Y. (Creator), Menzel, J. (Creator), Edenhofer, I. (Creator), Mezher, S. (Creator), Aypek, H. (Creator), Dumoulin, B. (Creator), Wu, H. (Creator), Hofmann, S. (Creator), Kretz, O. (Creator), Wanner, N. (Creator), Tomas, N. M. (Creator), Krasemann, S. (Creator), Glatzel, M. (Creator), Kuppe, C. (Creator), Kramann, R. (Creator), Banjanin, B. (Creator), Schneider, R. (Creator), Urbschat, C. (Creator), Arck, P. (Creator), Gagliani, N. (Creator), van Zandvoort, M. (Creator), Wiech, T. (Creator), Grahammer, F. (Creator), Sáez, P. J. (Creator), Wong, M. N. (Creator), Bonn, S. (Creator) & Puelles, V. G. (Creator), 10 Apr 2023


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