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Updated: Mar 2, 2026
Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions
Published on: June 24, 2013
Find your way with X-Ray: Using microCT to correlate in vivo imaging with 3D electron microscopy
Matthia A Karreman1, Bernhard Ruthensteiner2, Luc Mercier3
1European Molecular Biology Laboratory, Heidelberg, Germany.
Abstract:
Combining in vivo imaging with electron microscopy (EM) uniquely allows monitoring rare and critical events in living tissue, followed by their high-resolution visualization in their native context. A major hurdle, however, is to keep track of the region of interest (ROI) when moving from intravital microscopy (IVM) to EM. Here, we present a workflow that relies on correlating IVM and microscopic X-ray computed tomography to predict the position of the ROI inside the EM-processed sample. The ROI can then be accurately and quickly targeted using ultramicrotomy and imaged using EM. We outline how this procedure is used to retrieve and image tumor cells arrested in the vasculature of the mouse brain.
Insights
This study introduces a new workflow combining live imaging and electron microscopy to visualize rare cellular events. The method accurately tracks regions of interest, enabling high-resolution imaging of tumor cells in mouse brains.
Area of Science:
- Biological imaging
- Electron microscopy
- Cancer research
Background:
- Correlating live imaging with electron microscopy (EM) allows visualization of rare biological events in native tissue.
- A key challenge is relocating the region of interest (ROI) from live imaging to EM for detailed analysis.
Purpose of the Study:
- To develop and validate a workflow for accurately relocating ROIs from intravital microscopy (IVM) to EM.
- To enable high-resolution imaging of specific cellular events within their native tissue context.
Main Methods:
- Integration of intravital microscopy (IVM) with microscopic X-ray computed tomography.
- Development of a correlative workflow to predict ROI position in EM-processed samples.
- Utilizing ultramicrotomy for precise targeting and EM imaging of the ROI.
Main Results:
- Successfully correlated IVM data with EM imaging.
- Accurately identified and targeted ROIs within EM-processed samples.
- Visualized tumor cells arrested in the vasculature of the mouse brain at high resolution.
Conclusions:
- The presented workflow effectively bridges the gap between live imaging and EM.
- This method facilitates the study of rare and critical cellular events in their native environment.
- Enables precise retrieval and high-resolution imaging of specific biological targets, such as tumor cells in vivo.
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