Protocol for 3D-guided sectioning and deep cell phenotyping via light sheet imaging and 2D spatial multiplexing
Spatial analysis of cells and their microenvironment within tissues enhances our understanding of biological processes. Ideally, a broad range of biomolecules should be analyzed in large 3D tissue specimens at subcellular resolution. Here, we present a protocol to identify and extract target sections from previously cleared tissues. We describe steps for combining 3D light sheet imaging and subsequent 3D-guided deep cell phenotyping via multi-cyclic 2D microscopy.
Authors
- Kevin Bigott
- Marie-Catherine Tiveron (ORCID: https://orcid.org/0000-0003-3719-3494)
- Andreas Bosio (ORCID: https://orcid.org/0000-0001-9776-2535)
- Harold Cremer (ORCID: https://orcid.org/0000-0002-8673-5176)
- Melanie Jungblut (ORCID: https://orcid.org/0000-0001-9126-6610)
- Manuel Martinez-Osuna
- Christoph Herbel (ORCID: https://orcid.org/0000-0002-1933-3070)
- Victoria Schoppel
- Simon F. Bornemann
- Daniel Barleben
- Leon Osinski
Institutions
- Miltenyi Biotec (Germany) (DE)
- Institut de Biologie du Développement Marseille (FR)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-12-27
- DOI
- https://doi.org/10.1016/j.xpro.2025.104296
- Primary Topic
- Advanced Fluorescence Microscopy Techniques
- Type
- article
- Field-Weighted Citation Impact
- 0.00