VHUT-cryo-FIB
| Title | VHUT-cryo-FIB |
|---|---|
| Description | Cryo-electron tomography (cryo-ET) provides a promising approach to study intact structures of macromolecules in situ, but the efficient preparation of high-quality cryosections represents a bottleneck. Although cryo-focused ion beam (cryo-FIB) milling has emerged for large and flat cryo-lamella preparation, its application to tissue specimens remains challenging. Here, we report an integrated workflow, VHUT-cryo-FIB, for efficiently preparing frozen hydrated tissue lamella that can be readily used in subsequent cryo-ET studies. The workflow includes vibratome slicing, high-pressure freezing, ultramicrotome cryo-trimming and cryo-FIB milling. Two strategies were developed for loading cryo-lamella via a side-entry cryo-holder or an FEI AutoGrid. The workflow was validated by using various tissue specimens, including rat skeletal muscle, rat liver and spinach leaf specimens, and in situ structures of ribosomes were obtained at nanometer resolution from the spinach and liver samples. |
| Organism | Spinacia oleracea |
| Data Type | Other Type of Image Data |
| Data Accessibility | Open-access |
| BioProject | PRJCA016643 |
| Release Date | 2023-06-05 |
| Submitter | Shuoguo Li (lishuoguo@ibp.ac.cn) |
| Organization | Institute of Biophysics, Chinese Academy of Sciences |
| Submission Date | 2023-04-28 |
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| File ID | File Title | Number/Samples | File Type | File Size | File Suffix | Download Times | Download |
|---|---|---|---|---|---|---|---|
| OMIX003894-01 | liver20201028080914 | 3 | Other Type of Image Data | 19.23 GB | zip | 0 | |
| OMIX003894-02 | 20201012spinach05 | 1 | Other Type of Image Data | 7.68 GB | zip | 0 | |
| OMIX003894-05 | 27-Sep-2020 | 1 | Other Type of Image Data | 9.26 GB | zip | 0 | |
| OMIX003894-06 | VHUT-cryo-FIBworkflowandesign | 1 | Other Type of Image Data | 46.95 MB | zip | 0 | |
| OMIX003894-07 | liver20201108080910 | 3 | Other Type of Image Data | 18.64 GB | zip | 0 | |
| OMIX003894-08 | 20200927pinach14 | 1 | Other Type of Image Data | 10.26 GB | zip | 0 | |
| OMIX003894-09 | liver20201018 | 1 | Other Type of Image Data | 19.39 GB | zip | 0 | |
| OMIX003894-10 | 20201012spinach09 | 1 | Other Type of Image Data | 7.68 GB | zip | 0 | |
| OMIX003894-11 | 20200927pinach16 | 1 | Other Type of Image Data | 10.24 GB | zip | 0 | |
| OMIX003894-12 | VHUT-cryo-FIBprotocol | 1 | Other Type of Image Data | 2.06 MB | zip | 0 | |
| OMIX003894-13 | liver202010281516 | 2 | Other Type of Image Data | 13.51 GB | zip | 0 | |
| OMIX003894-14 | 20200927pinach15 | 1 | Other Type of Image Data | 10.28 GB | zip | 0 | |
| OMIX003894-15 | liver202011081112 | 1 | Other Type of Image Data | 12.69 GB | zip | 0 | |
| OMIX003894-16 | resultsofVHUTcryoFIB | 7 | Other Type of Image Data | 48.4 MB | zip | 0 |