Expanded vacuum-stable gels for multiplexed high-resolution spatial histopathology

Yunhao Bai, Bokai Zhu, John-Paul Oliveria, Bryan J. Cannon, Dorien Feyaerts, Marc Bosse, Kausalia Vijayaragavan, Noah F. Greenwald, Darci Phillips, Christian M. Schürch, Samuel M. Naik, Edward A. Ganio, Brice Gaudilliere, Scott J. Rodig, Michael B. Miller, Michael Angelo, Sean C. Bendall, Xavier Rovira-Clavé (), Garry P. Nolan () and Sizun Jiang ()
Additional contact information
Yunhao Bai: Stanford University
Bokai Zhu: Stanford University
John-Paul Oliveria: Genentech, Inc.
Bryan J. Cannon: Stanford University
Dorien Feyaerts: Stanford University
Marc Bosse: Stanford University
Kausalia Vijayaragavan: Stanford University
Noah F. Greenwald: Stanford University
Darci Phillips: Stanford University
Christian M. Schürch: Stanford University
Samuel M. Naik: Harvard Medical School
Edward A. Ganio: Stanford University
Brice Gaudilliere: Stanford University
Scott J. Rodig: Harvard Medical School
Michael B. Miller: Harvard Medical School
Michael Angelo: Stanford University
Sean C. Bendall: Stanford University
Xavier Rovira-Clavé: Stanford University
Garry P. Nolan: Stanford University
Sizun Jiang: Stanford University

Nature Communications, 2023, vol. 14, issue 1, 1-18

Abstract: Abstract Cellular organization and functions encompass multiple scales in vivo. Emerging high-plex imaging technologies are limited in resolving subcellular biomolecular features. Expansion Microscopy (ExM) and related techniques physically expand samples for enhanced spatial resolution, but are challenging to be combined with high-plex imaging technologies to enable integrative multiscaled tissue biology insights. Here, we introduce Expand and comPRESS hydrOgels (ExPRESSO), an ExM framework that allows high-plex protein staining, physical expansion, and removal of water, while retaining the lateral tissue expansion. We demonstrate ExPRESSO imaging of archival clinical tissue samples on Multiplexed Ion Beam Imaging and Imaging Mass Cytometry platforms, with detection capabilities of > 40 markers. Application of ExPRESSO on archival human lymphoid and brain tissues resolved tissue architecture at the subcellular level, particularly that of the blood-brain barrier. ExPRESSO hence provides a platform for extending the analysis compatibility of hydrogel-expanded biospecimens to mass spectrometry, with minimal modifications to protocols and instrumentation.

Date: 2023
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DOI: 10.1038/s41467-023-39616-w

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