| 1 | Nuclear peripheral chromatin-lamin B1 interaction is required for global integrity of chromatin architecture and dynamics in human cells显示文摘The eukaryotic genome is folded into higher-order conformation accompanied with constrained dynamics for coordinated genome functions.However,the molecular machinery underlying these hierarchically organized three-dimensional(3D)chromatin architecture and dynamics remains poorly understood.Here by combining imaging and sequencing,we studied the role of lamin B1 in chromatin architecture and dynamics.We found that lamin B1 depletion leads to detachment of lamina-associated domains(LADs)from the nuclear periphery accompanied with global chromatin redistribution and decompaction.Consequently,the interchromosomal as well as inter-compartment interactions are increased,but the structure of topologically associating domains(TADs)is not affected.Using live-cell genomic loci tracking,we further proved that depletion of lamin B1 leads to increased chromatin dynamics,owing to chromatin decompaction and redistribution toward nucleoplasm.Taken together,our data suggest that lamin B1 and chromatin interactions at the nuclear periphery promote LAD maintenance,chromatin compaction,genomic compartmentalization into chromosome territories and A/B compartments and confine chromatin dynamics,supporting their crucial roles in chromatin higher-order structure and chromatin dynamics. | Lei Chang Mengfan Li Shipeng Shao Chen Li Shanshan Ai Boxin Xue Yingping Hou Yiwen Zhang Ruifeng Li Xiaoying Fan Aibin He Cheng Li Yujie Sun | 2022 | Protein & Cell2022，13，4： | 0 |
| 2 | PN-ImTLSM facilitates high-throughput low background single-molecule localization microscopy deep in the cell显示文摘When imaging the nucleus structure of a cell,the out-of-focus fluorescence acts as background and hinders the detection of weak signals.Light-sheet fluorescence microscopy(LSFM)is a wide-field imaging approach which has the best of both background removal and imaging speed.However,the commonly adopted orthogonal excitation/detection scheme is hard to be applied to single-cell imaging due to steric hindrance.For LSFMs capable of high spatiotemporal single-cell imaging,the complex instrument design and operation largely limit their throughput of data collection.Here,we propose an approach for high-throughput background-free fluorescence imaging of single cells facilitated by the Im-mersion Tilted Light Sheet Microscopy(ImTLSM).ImTLSM is based on a light-sheet projected off the optical axis of a water immersion objective.With the illumination objective and the detection objective placed opposingly,ImTLSM can rapidly patrol and optically section multiple individual cells while maintaining single-molecule detection sensitivity and resolution.Further,the simplicity and robustness of ImTLSM in operation and maintenance enables high-throughput image collection to establish background removal datasets for deep learning.Using a deep learning model to train the mapping from epi-illumination images to ImTLSM illumination images,namely PN-ImTLSM,we demonstrated crossmodality fluorescence imaging,transforming the epi-illumination image to approach the background removal performance obtained with ImTLSM.We demonstrated that PN-ImTLSM can be generalized to large-field homogeneous illumination imaging,thereby further improving the imaging throughput.In addition,compared to commonly used background removal methods,PN-ImTLSM showed much better performance for areas where the background intensity changes sharply in space,facilitating high-density single-molecule localization microscopy.In summary,PN-ImTLSM paves the way for background-free fluorescence imaging on ordinary inverted microscopes. | Boxin Xue Caiwei Zhou YizhiQin Yongzheng Li Yuao Sun Lei Chang Shipeng Shao Yongliang Li Mengling Zhang Chaoying Sun Renxi He Qian Peter Su Yujie Sun | 2021 | Biophysics Reports2021，7，4： | 0 |
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