Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, UK.
Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15260, USA.
Structure. 2021 Jan 7;29(1):82-87.e3. doi: 10.1016/j.str.2020.10.003. Epub 2020 Oct 22.
The advancement of serial cryoFIB/SEM offers an opportunity to study large volumes of near-native, fully hydrated frozen cells and tissues at voxel sizes of 10 nm and below. We explored this capability for pathologic characterization of vitrified human patient cells by developing and optimizing a serial cryoFIB/SEM volume imaging workflow. We demonstrate profound disruption of subcellular architecture in primary fibroblasts from a Leigh syndrome patient harboring a disease-causing mutation in USMG5 protein responsible for impaired mitochondrial energy production.
连续冷冻断裂/扫描电镜的进步为我们提供了一个机会，使我们可以在 10nm 及以下的体素大小下，对近天然、完全水合的冷冻细胞和组织的大体积进行研究。我们通过开发和优化连续冷冻断裂/扫描电镜体积成像工作流程，探索了这种能力在病理解剖特征分析中的应用，对一位患有 Leigh 综合征的患者的人源细胞进行了冷冻成像，该患者携带有 USMG5 蛋白的致病突变，导致线粒体能量产生受损。
