Multi-Scale 3D Cryo-Correlative Microscopy for Vitrified Cells
Multi-Scale 3D Cryo-Correlative Microscopy for Vitrified Cells
复制标题
玻璃化细胞的多尺度 3D 冷冻相关显微镜
DOI：
10.1101/2020.05.21.107771
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
W. Chiu
中科院分区：
文献类型：
--
作者：
Gong;P. Mitchell;Jesús G. Galaz;Corey W. Hecksel;E. Sontag;V. Gangadharan;J. Marshman;D. Mankus;M. Bisher;Abigail K. R. Lytton;J. Frydman;K. Czymmek;W. Chiu
Three-dimensional (3D) visualization of vitrified cells can uncover structures of subcellular complexes without chemical fixation or staining. Here, we present a pipeline integrating three imaging modalities to visualize the same specimen at cryogenic temperature at different scales: cryo-fluorescence confocal microscopy, volume cryo-focused ion beam scanning electron microscopy, and transmission cryo-electron tomography. Our proof-of-concept benchmark revealed the 3D distribution of organelles and subcellular structures in whole heat-shocked yeast cells, including the ultrastructure of protein inclusions that recruit fluorescently-labelled chaperone Hsp104. Since our workflow efficiently integrates imaging at three different scales and can be applied to other types of cells, it could be used for large-scale phenotypic studies of frozen-hydrated specimens in a variety of healthy and diseased conditions with and without treatments.
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