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-Structure paper
タイトル | Multi-scale 3D Cryo-Correlative Microscopy for Vitrified Cells. |
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ジャーナル・号・ページ | Structure, Vol. 28, Issue 11, Page 1231-11237.e3, Year 2020 |
掲載日 | 2020年11月3日 |
著者 | Gong-Her Wu / Patrick G Mitchell / Jesus G Galaz-Montoya / Corey W Hecksel / Emily M Sontag / Vimal Gangadharan / Jeffrey Marshman / David Mankus / Margaret E Bisher / Abigail K R Lytton-Jean / Judith Frydman / Kirk Czymmek / Wah Chiu / |
PubMed 要旨 | 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 ...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-labeled 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. |
リンク | Structure / PubMed:32814034 / PubMed Central |
手法 | EM (サブトモグラム平均) / EM (トモグラフィー) |
解像度 | 21.0 Å |
構造データ | EMDB-21952: EMDB-21953: |
由来 |
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