Array tomography (AT) is a high-resolution imaging method to resolve fine details at the organelle level and has the advantage that it can provide 3D volumes to show the tissue context. AT can be carried out in a correlative way, combing light and electron microscopy (LM, EM) techniques. However, the correlation between modalities can be a challenge and delineating specific regions of interest in consecutive sections can be time-consuming. Integrated light and electron microscopes (iLEMs) offer the possibility to provide well-correlated images and may pose an ideal solution for correlative AT. Here, we report a workflow to automate navigation between regions of interest. We use a targeted approach that allows imaging specific tissue features, like organelles, cell processes, and nuclei at different scales to enable fast, directly correlated in situ AT using an integrated light and electron microscope (iLEM-AT). Our workflow is based on the detection of section boundaries on an initial transmitted light acquisition that serves as a reference space to compensate for changes in shape between sections, and we apply a stepwise refinement of localizations as the magnification increases from LM to EM. With minimal user interaction, this enables autonomous and speedy acquisition of regions containing cells and cellular organelles of interest correlated across different magnifications for LM and EM modalities, providing a more efficient way to obtain 3D images. We provide a proof of concept of our approach and the developed software tools using both Golgi neuronal impregnation staining and fluorescently labeled protein condensates in cells. Our method facilitates tracing and reconstructing cellular structures over multiple sections, is targeted at high resolution ILEMs, and can be integrated into existing devices, both commercial and custom-built systems.

中文翻译：

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阵列断层扫描中串行感兴趣区域的简化采集和关联工作流程

阵列断层扫描 (AT) 是一种高分辨率成像方法，可在细胞器水平解析精细细节，并具有可以提供 3D 体积以显示组织背景的优势。AT 可以以相关的方式进行，结合光学和电子显微镜（LM，EM）技术。然而，模态之间的相关性可能是一个挑战，并且在连续部分中描绘特定的感兴趣区域可能很耗时。集成光学和电子显微镜 (iLEM) 提供了提供良好相关图像的可能性，并可能为相关 AT 提供理想的解决方案。在这里，我们报告了一个在感兴趣区域之间自动导航的工作流程。我们使用有针对性的方法，允许对特定组织特征（如不同尺度的细胞器、细胞过程和细胞核）进行成像，以实现快速、使用集成光学和电子显微镜 (iLEM-AT) 直接相关的原位 AT。我们的工作流程基于在初始透射光采集中检测截面边界，该采集用作参考空间以补偿截面之间的形状变化，并且随着放大率从 LM 增加到 EM，我们对定位进行逐步细化。通过最少的用户交互，这可以自主和快速地获取包含在 LM 和 EM 模式的不同放大倍率下相关的感兴趣的细胞和细胞器的区域，从而提供一种更有效的方式来获取 3D 图像。我们使用高尔基神经元浸渍染色和细胞中荧光标记的蛋白质凝聚物提供了我们的方法和开发的软件工具的概念证明。