TY - JOUR T1 - Light and electron microscopy continuum-resolution imaging of 3D cell cultures JF - bioRxiv DO - 10.1101/2021.07.02.450855 SP - 2021.07.02.450855 AU - Edoardo D’Imprima AU - Marta Garcia Montero AU - Sylwia Gawrzak AU - Paolo Ronchi AU - Ievgeniia Zagoriy AU - Yannick Schwab AU - Martin Jechlinger AU - Julia Mahamid Y1 - 2022/01/01 UR - http://biorxiv.org/content/early/2022/12/13/2021.07.02.450855.abstract N2 - 3D cell cultures, in particular organoids, are emerging models to investigate healthy or diseased tissues. Understanding the complex cellular sociology in organoids requires integration of imaging modalities across spatial and temporal scales. We present a multi-scale imaging approach that traverses millimeter-scale live-cell light microscopy to nano-scale volume electron microscopy by performing 3D cell cultures in a single carrier amenable to all imaging steps. This allows to follow organoids growth, probe their morphology with fluorescent markers, identify areas of interest and analyze their 3D ultrastructure. We demonstrate this workflow on mouse and human 3D cultures, and use automated image segmentation to annotate and quantitatively analyze subcellular structures in patient-derived colorectal cancer organoids. Our analyses reveal local organization of diffraction-limited cell junctions in compact and polarized epithelia. The continuum resolution imaging pipeline is thus suited to foster basic and translational organoid research by simultaneously exploiting the advantages of light and electron microscopy.HighlightsEstablishment of 3D cell cultures in sample carriers directly amenable to high-pressure freezing (HPF)3D cell cultures in HPF carriers allow drug treatment and live-cell imagingMulti-scale imaging of 3D cultures from live-cell light microscopy to volume electron microscopyEstablishments of HPF conditions for mouse and patient-derived organoidsDeep-learning automatic segmentation of ultrastructural detail and quantitative data-mining reveal different subcellular organization associated with epithelium polarityCompeting Interest StatementThe authors have declared no competing interest. ER -