Abstract：
Three-dimensional structured illumination microscopy (3D-SIM) and fluorescence in situ hybridization on three-dimensional preserved cells (3D-FISH) have proven to be robust and efficient methodologies for analyzing nuclear architecture and profiling the genome\'s topological features. These methods have allowed the simultaneous visualization and evaluation of several target structures at super-resolution. In this chapter, we focus on the application of 3D-SIM for the visualization of 3D-FISH preparations of chromosomes in interphase, known as Chromosome Territories (CTs). We provide a workflow and detailed guidelines for sample preparation, image acquisition, and image analysis to obtain quantitative measurements for profiling chromosome topological features. In parallel, we address a practical example of these protocols in the profiling of CTs 9 and 22 involved in the translocation t(9;22) in Chronic Myeloid Leukemia (CML). The profiling of chromosome topological features described in this chapter allowed us to characterize a large-scale topological disruption of CTs 9 and 22 that correlates directly with patients\' response to treatment and as a possible potential change in the inheritance systems. These findings open new insights into how the genome structure is associated with the response to cancer treatments, highlighting the importance of microscopy in analyzing the topological features of the genome.
摘要：
三维结构化照明显微镜（3D-SIM）和三维保存细胞的荧光原位杂交（3D-FISH）已被证明是分析核结构和分析基因组拓扑特征的强大而有效的方法。这些方法允许以超分辨率同时可视化和评估多个目标结构。在这一章中,我们专注于3D-SIM在相间染色体的3D-FISH制剂可视化中的应用，称为染色体区域（CT）。我们提供样品制备的工作流程和详细指南，图像采集,和图像分析，以获得定量测量，以分析染色体拓扑特征。并行,我们在慢性髓系白血病(CML)中涉及t(9;22)易位的CT9和22的分析中,讨论了这些方案的一个实际例子.本章中描述的染色体拓扑特征的分析使我们能够描述与患者对治疗的反应直接相关的CT9和22的大规模拓扑破坏，并作为遗传系统中可能的潜在变化。这些发现为基因组结构如何与癌症治疗反应相关开辟了新的见解。强调显微镜在分析基因组拓扑特征中的重要性。
