title: Fast and precise targeting of single tumor cells in vivo by multimodal correlative microscopy.
abstract: Intravital microscopy provides dynamic understanding of multiple cell biological processes, but its limited resolution has so far precluded structural analysis. Because it is difficult to capture rare and transient events, only a few attempts have been made to observe specific developmental and pathological processes in animal models using electron microscopy. The multimodal correlative approach that we propose here combines intravital microscopy, microscopic X-ray computed tomography and three-dimensional electron microscopy. It enables a rapid (c.a. 2 weeks) and accurate (<5 µm) correlation of functional imaging to ultrastructural analysis of single cells in a relevant context. We demonstrate the power of our approach by capturing single tumor cells in the vasculature of the cerebral cortex and in subcutaneous tumors, providing unique insights into metastatic events. Providing a significantly improved throughput, our workflow enables multiple sampling, a prerequisite for making correlative imaging a relevant tool to study cell biology in vivo. Owing to the versatility of this workflow, we envision broad applications in various fields of biological research, such as cancer or developmental biology.
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg 69117, Germany.
authors.forename: Matthia A
authors.initials: MA
authors.lastname: Karreman
authors.affiliation: MN3T, Inserm U1109, Strasbourg 67200, France Université de Strasbourg, Strasbourg 67000, France LabEx Medalis, Université de Strasbourg, Strasbourg 67000, France Fédération de Médecine Translationnelle de Strasbourg (FMTS), Université de Strasbourg, Strasbourg 67000, France.
authors.forename: Luc
authors.initials: L
authors.lastname: Mercier
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg 69117, Germany.
authors.forename: Nicole L
authors.initials: NL
authors.lastname: Schieber
authors.affiliation: Department of Neurooncology, University Hospital Heidelberg, Heidelberg 69120, Germany Clinical Cooperation Unit Neurooncology, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany.
authors.forename: Gergely
authors.initials: G
authors.lastname: Solecki
authors.affiliation: MN3T, Inserm U1109, Strasbourg 67200, France Université de Strasbourg, Strasbourg 67000, France LabEx Medalis, Université de Strasbourg, Strasbourg 67000, France Fédération de Médecine Translationnelle de Strasbourg (FMTS), Université de Strasbourg, Strasbourg 67000, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Allio
authors.affiliation: Department of Neurooncology, University Hospital Heidelberg, Heidelberg 69120, Germany Clinical Cooperation Unit Neurooncology, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Winkler
authors.affiliation: Evertebrata Varia, Zoologische Staatssammlung München, Munich 81247, Germany.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Ruthensteiner
authors.affiliation: MN3T, Inserm U1109, Strasbourg 67200, France Université de Strasbourg, Strasbourg 67000, France LabEx Medalis, Université de Strasbourg, Strasbourg 67000, France Fédération de Médecine Translationnelle de Strasbourg (FMTS), Université de Strasbourg, Strasbourg 67000, France jacky.goetz@inserm.fr schwab@embl.de.
authors.forename: Jacky G
authors.initials: JG
authors.lastname: Goetz
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg 69117, Germany jacky.goetz@inserm.fr schwab@embl.de.
authors.forename: Yannick
authors.initials: Y
authors.lastname: Schwab
mesh_terms: D000818:Animals; D001921:Brain / Q000098:blood supply / Q000473:pathology; D001932:Brain Neoplasms / Q000000981:diagnostic imaging / Q000556:secondary; D045744:Cell Line, Tumor; D058948:Cell Tracking / Q000379:methods*; D005260:Female; D000069416:Intravital Microscopy; D008819:Mice, Nude; D008855:Microscopy, Electron, Scanning; D009368:Neoplasm Transplantation; D059016:Tumor Microenvironment; D055114:X-Ray Microtomography
keywords: Correlative microscopy; Electron microscopy; Extravasation; Intravital imaging; Invasion; Metastasis