| Accession |
PRJCA015542 |
| Title |
Cryo-ET data of human centrosome in situ |
| Relevance |
Medical |
| Data types |
Imaging data
|
| Organisms |
Homo sapiens
Homo
Human betaherpesvirus 6
|
| Description |
Cryo-electron tomography (cryo-ET) has become a powerful approach to study the high-resolution structure of cellular macromolecular machines in situ. However, the current correlative cryo-fluorescence and electron microscopy lacks sufficient accuracy and efficiency to precisely prepare cryo-lamellae of target locations for subsequent cryo-ET study. Here, we developed a precise cryogenic fabrication system, ELI-TriScope, by setting up an electron beam, a light beam and an ion beam at the same focal point to achieve accurate and efficient preparation of a target cryo-lamella. ELI-TriScope was developed starting from a commercial dual-beam scanning electron microscope by incorporating a cryo-holder-based transfer system and embedding an optical imaging system just underneath the vitrified specimen. Cryo-focused ion beam milling can be accurately navigated by monitoring the real-time fluorescence signal of the target molecule. Using ELI-TriScope, we prepared a batch of cryo-lamellae of HeLa cells targeting the centrosome, with a success rate of ~91%, and discovered new in situ structural features of the human centrosome by cryo-ET. |
| Sample scope |
Monoisolate |
| Release date |
2023-04-28 |
| Publication |
|
| Grants |
| Agency |
program |
Grant ID |
Grant title |
| Chinese Academy of Sciences (CAS)
|
Strategic Priority Research Program of Chinese Academy of Sciences (CAS)
|
XDB37000000
|
|
|
|
| Submitter |
Shuoguo
Li (lishuoguo@ibp.ac.cn)
|
| Organization |
Institute of Biophysics, Chinese Academy of Sciences |
| Submission date |
2023-03-14 |
