Randall Centre for Cell and Molecular Biophysics, King's College London, New Hunts House, Guys Campus, London SE1 1UL, UK.
Microscopy Innovation Centre, King's College London, Guys Campus, London SE1 1UL, UK.
J Cell Sci. 2024 Jan 15;137(2). doi: 10.1242/jcs.261389. Epub 2024 Jan 18.
The actin cytoskeleton plays a critical role in cell architecture and the control of fundamental processes including cell division, migration and survival. The dynamics and organisation of F-actin have been widely studied in a breadth of cell types on classical two-dimensional (2D) surfaces. Recent advances in optical microscopy have enabled interrogation of these cytoskeletal networks in cells within three-dimensional (3D) scaffolds, tissues and in vivo. Emerging studies indicate that the dimensionality experienced by cells has a profound impact on the structure and function of the cytoskeleton, with cells in 3D environments exhibiting cytoskeletal arrangements that differ to cells in 2D environments. However, the addition of a third (and fourth, with time) dimension leads to challenges in sample preparation, imaging and analysis, necessitating additional considerations to achieve the required signal-to-noise ratio and spatial and temporal resolution. Here, we summarise the current tools for imaging actin in a 3D context and highlight examples of the importance of this in understanding cytoskeletal biology and the challenges and opportunities in this domain.
肌动蛋白细胞骨架在细胞结构和基本过程（包括细胞分裂、迁移和存活）的控制中起着关键作用。在经典二维（2D）表面上，广泛研究了 F-肌动蛋白的动力学和组织。光学显微镜的最新进展使人们能够在三维（3D）支架、组织和体内的细胞中研究这些细胞骨架网络。新兴研究表明，细胞所经历的维度对细胞骨架的结构和功能有深远的影响，与 2D 环境中的细胞相比，3D 环境中的细胞表现出不同的细胞骨架排列。然而，增加第三个（以及第四个，随着时间的推移）维度会给样品制备、成像和分析带来挑战，需要额外的考虑来实现所需的信噪比和空间和时间分辨率。在这里，我们总结了当前在 3D 环境中成像肌动蛋白的工具，并强调了在理解细胞骨架生物学方面的重要性，以及在该领域的挑战和机遇。
