Zhejiang Provincial Key Laboratory of Biometrology and Inspection and Quarantine, College of Life Science, China , Jiliang University, Hangzhou, 310018, China.
Cell Tissue Res. 2024 Apr;396(1):1-18. doi: 10.1007/s00441-024-03878-7. Epub 2024 Feb 28.
Imaging technologies have played a pivotal role in advancing biological research by enabling visualization of biological structures and processes. While traditional electron microscopy (EM) produces two-dimensional images, emerging techniques now allow high-resolution three-dimensional (3D) characterization of specimens in situ, meeting growing needs in molecular and cellular biology. Combining transmission electron microscopy (TEM) with serial sectioning inaugurated 3D imaging, attracting biologists seeking to explore cell ultrastructure and driving advancement of 3D EM reconstruction. By comprehensively and precisely rendering internal structure and distribution, 3D TEM reconstruction provides unparalleled ultrastructural insights into cells and molecules, holding tremendous value for elucidating structure-function relationships and broadly propelling structural biology. Here, we first introduce the principle of 3D reconstruction of cells and tissues by classical approaches in TEM and then discuss modern technologies utilizing TEM and on new SEM-based as well as cryo-electron microscope (cryo-EM) techniques. 3D reconstruction techniques from serial sections, electron tomography (ET), and the recent single-particle analysis (SPA) are examined; the focused ion beam scanning electron microscopy (FIB-SEM), the serial block-face scanning electron microscopy (SBF-SEM), and automatic tape-collecting lathe ultramicrotome (ATUM-SEM) for 3D reconstruction of large volumes are discussed. Finally, we review the challenges and development prospects of these technologies in life science. It aims to provide an informative reference for biological researchers.
成像技术在推进生物学研究方面发挥了关键作用，使生物结构和过程的可视化成为可能。虽然传统的电子显微镜（EM）产生二维图像，但新兴技术现在允许对原位标本进行高分辨率的三维（3D）表征，满足了分子和细胞生物学不断增长的需求。透射电子显微镜（TEM）与连续切片的结合开创了 3D 成像，吸引了寻求探索细胞超微结构的生物学家，并推动了 3D EM 重建的发展。通过全面准确地呈现内部结构和分布，3D TEM 重建为细胞和分子提供了无与伦比的超微结构见解，对于阐明结构-功能关系和广泛推动结构生物学具有巨大价值。在这里，我们首先介绍 TEM 中经典方法重建细胞和组织的 3D 原理，然后讨论利用 TEM 和新的基于 SEM 以及冷冻电子显微镜（cryo-EM）技术的现代技术。检查了从连续切片、电子断层扫描（ET）和最近的单颗粒分析（SPA）的 3D 重建技术；讨论了用于大体积 3D 重建的聚焦离子束扫描电子显微镜（FIB-SEM）、连续块面扫描电子显微镜（SBF-SEM）和自动纸带收集车床超微切片机（ATUM-SEM）。最后，我们回顾了这些技术在生命科学中的挑战和发展前景。旨在为生物研究人员提供有价值的参考。
