TY - JOUR
T1 - Expansion-enhanced super-resolution radial fluctuations enable nanoscale molecular profiling of pathology specimens
AU - Kylies, Dominik
AU - Zimmermann, Marina
AU - Haas, Fabian
AU - Schwerk, Maria
AU - Kuehl, Malte
AU - Brehler, Michael
AU - Czogalla, Jan
AU - Hernandez, Lola C.
AU - Konczalla, Leonie
AU - Okabayashi, Yusuke
AU - Menzel, Julia
AU - Edenhofer, Ilka
AU - Mezher, Sam
AU - Aypek, Hande
AU - Dumoulin, Bernhard
AU - Wu, Hui
AU - Hofmann, Smilla
AU - Kretz, Oliver
AU - Wanner, Nicola
AU - Tomas, Nicola M.
AU - Krasemann, Susanne
AU - Glatzel, Markus
AU - Kuppe, Christoph
AU - Kramann, Rafael
AU - Banjanin, Bella
AU - Schneider, Rebekka K.
AU - Urbschat, Christopher
AU - Arck, Petra
AU - Gagliani, Nicola
AU - van Zandvoort, Marc
AU - Wiech, Thorsten
AU - Grahammer, Florian
AU - Saez, Pablo J.
AU - Wong, Milagros N.
AU - Bonn, Stefan
AU - Huber, Tobias B.
AU - Puelles, Victor G.
PY - 2023/4/1
Y1 - 2023/4/1
N2 - 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.
AB - 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.
KW - MICROSCOPY
KW - MEGAKARYOCYTE
KW - PLATELET
KW - DISEASE
U2 - 10.1038/s41565-023-01328-z
DO - 10.1038/s41565-023-01328-z
M3 - Article
C2 - 37037895
SN - 1748-3387
VL - 18
SP - 336
EP - 342
JO - Nature Nanotechnology
JF - Nature Nanotechnology
IS - 4
ER -