Unraveling Subcellular Ultrastructure with Cyclically Multiplexed Expansion Microscopy
Despite advances in fluorescence microscopy, spectral overlap and limited resolution hinder the dense mapping of the cellular ultrastructure. To overcome these challenges, we developed Cy-ExM, a high-plex imaging strategy that integrates optimized cryo-fixation for antigen preservation, expansion microscopy, and iterative immunofluorescence labeling. Using oblique plane microscopy, we perform three-dimensional super-resolution imaging of 20 biological targets encompassing the full cellular volume of individual mammalian cells.
Authors
- Daniel Stoddard (ORCID: https://orcid.org/0000-0001-9666-7923)
- Daniela Nicastro (ORCID: https://orcid.org/0000-0002-0122-7173)
- Reto Fiolka (ORCID: https://orcid.org/0000-0002-4636-5000)
- Seweryn Gałecki (ORCID: https://orcid.org/0000-0002-7020-0804)
- Felix Zhou (ORCID: https://orcid.org/0000-0003-4463-1165)
- Qionghua Shen (ORCID: https://orcid.org/0000-0002-2808-4408)
- Bingying Chen (ORCID: https://orcid.org/0000-0003-3518-6290)
- Bo-Jui Chang
- Kevin M. Dean
Institutions
- Silesian University of Technology (PL)
- Southwestern Medical Center (US)
- Southwestern Medical Center (US)
- The University of Texas Southwestern Medical Center (US)
Publication Details
- Journal
- bioRxiv (Cold Spring Harbor Laboratory)
- Published
- 2026-01-02
- DOI
- https://doi.org/10.64898/2026.01.01.697161
- Primary Topic
- Advanced Fluorescence Microscopy Techniques
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- University of Texas Southwestern Medical Center
- National Institutes of Health
- National Cancer Institute
- National Institute of General Medical Sciences
- National Institute of Biomedical Imaging and Bioengineering