Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana 47907, USA; email:
Institute for Quantitative Biomedicine, Rutgers University, Piscataway, New Jersey 08854, USA.
Annu Rev Biomed Eng. 2020 Jun 4;22:155-184. doi: 10.1146/annurev-bioeng-060418-052203. Epub 2020 Apr 3.
Super-resolution microscopy techniques are versatile and powerful tools for visualizing organelle structures, interactions, and protein functions in biomedical research. However, whole-cell and tissue specimens challenge the achievable resolution and depth of nanoscopy methods. We focus on three-dimensional single-molecule localization microscopy and review some of the major roadblocks and developing solutions to resolving thick volumes of cells and tissues at the nanoscale in three dimensions. These challenges include background fluorescence, system- and sample-induced aberrations, and information carried by photons, as well as drift correction, volume reconstruction, and photobleaching mitigation. We also highlight examples of innovations that have demonstrated significant breakthroughs in addressing the abovementioned challenges together with their core concepts as well as their trade-offs.
超分辨率显微镜技术是生物医学研究中可视化细胞器结构、相互作用和蛋白质功能的通用且强大的工具。然而，全细胞和组织标本对纳米显微镜方法的可实现分辨率和深度提出了挑战。我们专注于三维单分子定位显微镜，并综述了一些主要的障碍和正在开发的解决方案，以在三维尺度上解析厚细胞和组织的纳米尺度。这些挑战包括背景荧光、系统和样品诱导的像差以及光子携带的信息，以及漂移校正、体积重建和光漂白缓解。我们还强调了一些创新的例子，这些例子展示了在解决上述挑战方面的重大突破，以及它们的核心概念及其权衡。
