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Integrated light and electron microscopy continuum resolution imaging of 3D cell cultures

By Edoardo Dimprima, Marta Garcia Montero, Sylwia Gawrzak, Paolo Ronchi, Ievgeniia Zagoriy, Yannick Schwab, Martin Jechlinger, Julia Mahamid

Posted 03 Jul 2021
bioRxiv DOI: 10.1101/2021.07.02.450855

3D cell cultures, in particular organoids, recapitulate tissue architectures and are emerging model systems to investigate healthy or diseased tissues and organs. Understanding the complex cellular sociology of organoids requires the integration of diverse imaging modalities covering different spatial and temporal resolutions. Here, we present an integrated multi-scale 3D imaging approach that traverses from millimeter-scale live-cell light microscopy to nanoscale volume electron microscopy. Performing the 3D cell culture in a single carrier amenable to all imaging steps provides a continuum resolution view of the same organoid. Our approach allows us to follow organoid growth, probe their morphology with fluorescent markers, identify cells of interest and analyze their ultrastructure. We demonstrate this workflow on mouse and human derived 3D cell cultures. The continuum resolution imaging pipeline is thus suited to foster both basic and translational organoid research by simultaneously exploiting the advantages of light and electron microscopy.

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