Super-resolution microscopy, and specifically single-molecule localization microscopy (SMLM), is becoming a transformative technology for cell biology, as it allows the study of cellular structures with nanometer resolution. Here, we review a wide range of data analyses approaches for SMLM that extract quantitative information about the distribution, size, shape, spatial organization, and stoichiometry of macromolecular complexes to guide biological interpretation. We present a case study using the nuclear pore complex as an example that highlights the power of combining complementary approaches by identifying its symmetry, ringlike structure, and protein copy number. In face of recent technical and computational advances, this review serves as a guideline for selecting appropriate analysis tools and controls to exploit the potential of SMLM for a wide range of biological questions.

超分辨率显微镜，特别是单分子定位显微镜 (SMLM)，正在成为细胞生物学的一项变革性技术，因为它允许以纳米分辨率研究细胞结构。在这里，我们回顾了广泛的 SMLM 数据分析方法，这些方法提取关于大分子复合物的分布、大小、形状、空间组织和化学计量的定量信息，以指导生物学解释。我们展示了一个使用核孔复合体的案例研究，通过识别其对称性、环状结构和蛋白质拷贝数，突出了结合互补方法的力量。面对最近的技术和计算进步，