3D细胞培养，特别是器官样结构，是研究健康或患病组织的新模型。了解器官样结构内复杂的细胞社会需要在空间和时间尺度上整合成像模式。我们提出了一种多尺度成像方法，通过在单一载体中进行3D细胞培养，穿越毫米级现场活细胞光显微镜到纳米级三维体积电子显微镜，适合于所有成像步骤。这使得我们可以跟踪器官样结构的生长，用荧光标记物研究其形态，确定感兴趣的区域，并分析其三维超结构。我们在小鼠和人类3D培养上展示了这一工作流程，并使用自动图像分割注释和定量分析了患者来源的结直肠癌器官样结构中的亚细胞结构。我们的分析确定了紧密极性上皮组织中衍射极限细胞连接的局部组织。这种连续分辨率的成像管线适合于同时利用光学和电子显微镜的优势，促进基础和转化器官样结构研究。版权所有©2023作者。Elsevier Inc.保留所有权利。

3D cell cultures, in particular organoids, are emerging models in the investigation of 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 nanometer-scale volume electron microscopy by performing 3D cell cultures in a single carrier that is amenable to all imaging steps. This allows for following organoids' growth, probing their morphology with fluorescent markers, identifying areas of interest, and analyzing 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 identify local organization of diffraction-limited cell junctions in compact and polarized epithelia. The continuum-resolution imaging pipeline is thus suited to fostering basic and translational organoid research by simultaneously exploiting the advantages of light and electron microscopy.Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.