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Updated: Mar 28, 2026
Intravital Microscopy of Tumor-associated Vasculature Using Advanced Dorsal Skinfold Window Chambers on Transgenic Fluorescent Mice
Published on: January 19, 2018
Fast and precise targeting of single tumor cells in vivo by multimodal correlative microscopy
Matthia A Karreman1, Luc Mercier2, Nicole L Schieber1
1Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg 69117, Germany.
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.
Insights
This study introduces a novel multimodal imaging workflow combining intravital microscopy, X-ray computed tomography, and 3D electron microscopy. This method enables rapid, high-resolution ultrastructural analysis of dynamic cellular processes in vivo, offering new insights into cancer metastasis.
Area of Science:
- Cell Biology
- Microscopy
- Biophysics
Background:
- Intravital microscopy offers dynamic insights into cellular processes but lacks resolution for structural analysis.
- Observing rare, transient events in vivo using electron microscopy is challenging and limited.
- Current methods struggle to bridge functional imaging with detailed ultrastructural examination in a biological context.
Purpose of the Study:
- To develop a multimodal correlative imaging approach integrating intravital microscopy with advanced electron microscopy techniques.
- To enable rapid and accurate correlation of live-cell dynamics with ultrastructural details in vivo.
- To overcome limitations in observing and analyzing rare cellular events and structures within their native environment.
Main Methods:
- Combining intravital microscopy for live imaging with micro-X-ray computed tomography and 3D electron microscopy.
- Implementing a workflow for rapid (approx. 2 weeks) and accurate (<5 µm) correlation of functional and structural data.
- Utilizing multiple sampling capabilities to enhance throughput for correlative imaging studies.
Main Results:
- Successfully captured single tumor cells in the cerebral cortex vasculature and subcutaneous tumors.
- Provided unique ultrastructural insights into metastatic events by correlating live imaging with high-resolution electron microscopy.
- Demonstrated a significantly improved throughput for correlative imaging, facilitating in vivo cell biology studies.
Conclusions:
- The proposed multimodal correlative approach bridges the gap between functional intravital microscopy and ultrastructural analysis.
- This workflow provides unprecedented insights into dynamic cellular processes, particularly metastatic events, in vivo.
- The versatility and efficiency of this method offer broad applications in cancer biology, developmental biology, and beyond.
