The eukaryotic cell nucleus is a central organelle whose architecture determines genome function at multiple levels. Deciphering nuclear organizing principles influencing cellular responses and identity is a timely challenge. Despite many similarities between plant and animal nuclei, plant nuclei present intriguing specificities. Complementary to molecular and biochemical approaches, 3D microscopy is indispensable for resolving nuclear architecture. However, novel solutions are required for capturing cell-specific, sub-nuclear and dynamic processes. We provide a pointer for utilising high-to-super-resolution microscopy and image processing to probe plant nuclear architecture in 3D at the best possible spatial and temporal resolution and at quantitative and cell-specific levels. High-end imaging and image-processing solutions allow the community now to transcend conventional practices and benefit from continuously improving approaches. These promise to deliver a comprehensive, 3D view of plant nuclear architecture and to capture spatial dynamics of the nuclear compartment in relation to cellular states and responses.

Abbreviations: 3D and 4D: Three and Four dimensional; AI: Artificial Intelligence; ant: antipodal nuclei (ant); CLSM: Confocal Laser Scanning Microscopy; CTs: Chromosome Territories; DL: Deep Learning; DLIm: Dynamic Live Imaging; ecn: egg nucleus; FACS: Fluorescence-Activated Cell Sorting; FISH: Fluorescent In Situ Hybridization; FP: Fluorescent Proteins (GFP, RFP, CFP, YFP, mCherry); FRAP: Fluorescence Recovery After Photobleaching; GPU: Graphics Processing Unit; KEEs: KNOT Engaged Elements; INTACT: Isolation of Nuclei TAgged in specific Cell Types; LADs: Lamin-Associated Domains; ML: Machine Learning; NA: Numerical Aperture; NADs: Nucleolar Associated Domains; PALM: Photo-Activated Localization Microscopy; Pixel: Picture element; pn: polar nuclei; PSF: Point Spread Function; RHF: Relative Heterochromatin Fraction; SIM: Structured Illumination Microscopy; SLIm: Static Live Imaging; SMC: Spore Mother Cell; SNR: Signal to Noise Ratio; SRM: Super-Resolution Microscopy; STED: STimulated Emission Depletion; STORM: STochastic Optical Reconstruction Microscopy; syn: synergid nuclei; TADs: Topologically Associating Domains; Voxel: Volumetric pixel

真核细胞核是中央细胞器，其结构决定了多个水平的基因组功能。破解影响细胞反应和身份的核组织原则是一个及时的挑战。尽管动植物核之间有许多相似之处，但动植物核仍表现出令人着迷的特异性。作为分子和生化方法的补充，3D显微镜对于解析核结构是必不可少的。但是，需要新颖的解决方案来捕获特定于细胞的，亚核和动态过程。我们提供了一个利用超高分辨率的显微技术和图像处理技术，以最佳的时空分辨率以及定量和细胞特异性水平，以3D模式探测植物核结构的指针。高端成像和图像处理解决方案使社区现在可以超越常规做法，并受益于不断改进的方法。这些有望提供植物核结构的全面3D视图，并捕获与细胞状态和响应有关的核区室空间动态。

缩略语：3D和4D：3维和4维；AI：人工智能；ant：对足核（ant）；CLSM：共聚焦激光扫描显微镜；CT：染色体区域；DL：深度学习；DLIm：动态实时成像；ecn：蛋核；FACS：荧光激活细胞分选；鱼：荧光原位杂交；FP：荧光蛋白（GFP，RFP，CFP，YFP，mCherry）；FRAP：光漂白后的荧光恢复；GPU：图形处理单元；KEEs：不参与元素；目的：分离在特定细胞类型中捕获的细胞核；LAD：Lamin关联域；ML：机器学习；NA：数值孔径；NADs：核仁相关域；PALM：光激活定位显微镜；像素：像素 pn：极核；PSF：点扩展功能；RHF：相对异染色质分数；SIM：结构照明显微镜；瘦：静态实时成像；SMC：孢子母细胞；SNR：信噪比；SRM：超分辨率显微镜；STED：估算的排放耗竭；风暴：随机光学重建显微镜；syn：协同核；TAD：拓扑关联域；体素：体积像素