State Key Laboratory of Protein and Plant Gene Research, Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, School of Life Sciences, Peking University, Beijing 100871, China.
School of Life Sciences, Peking University, Beijing 100871, China.
J Struct Biol. 2023 Sep;215(3):107971. doi: 10.1016/j.jsb.2023.107971. Epub 2023 May 16.
The structural studies of macromolecules in their physiological context, particularly in tissue, is constrained by the bottleneck of sample preparation. In this study, we present a practical pipeline for preparing multicellular samples for cryo-electron tomography. The pipeline comprises sample isolation, vitrification, and lift-out-based lamella preparation using commercially available instruments. We demonstrate the efficacy of our pipeline by visualizing pancreatic β cells from mouse islets at the molecular level. This pipeline enables the determination of the properties of insulin crystals in situ for the first time, using unperturbed samples.
在生理环境中对生物大分子进行结构研究，特别是在组织中，受到样品制备瓶颈的限制。在这项研究中，我们提出了一个用于制备用于冷冻电子断层扫描的多细胞样品的实用流程。该流程包括使用市售仪器进行样品分离、玻璃化和基于提起的薄片制备。我们通过在分子水平上可视化来自小鼠胰岛的胰腺β细胞来证明我们的流程的功效。该流程使我们能够首次使用未受干扰的样品原位确定胰岛素晶体的性质。
