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Title | A practical multicellular sample preparation pipeline broadens the application of in situ cryo-electron tomography. |
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Journal, issue, pages | J Struct Biol, Vol. 215, Issue 3, Page 107971, Year 2023 |
Publish date | May 16, 2023 |
Authors | Yichun Wu / Changdong Qin / Wenjing Du / Zhenxi Guo / Liangyi Chen / Qiang Guo / |
PubMed Abstract | The structural studies of macromolecules in their physiological context, particularly in tissue, is constrained by the bottleneck of sample preparation. In this study, we present a practical pipeline ...The structural studies of macromolecules in their physiological context, particularly in tissue, is constrained by the bottleneck of sample preparation. In this study, we present a practical pipeline for preparing multicellular samples for cryo-electron tomography. The pipeline comprises sample isolation, vitrification, and lift-out-based lamella preparation using commercially available instruments. We demonstrate the efficacy of our pipeline by visualizing pancreatic β cells from mouse islets at the molecular level. This pipeline enables the determination of the properties of insulin crystals in situ for the first time, using unperturbed samples. |
External links | J Struct Biol / PubMed:37201639 |
Methods | EM (tomography) |
Structure data | EMDB-35652: cryo-electron tomogram from mouse islets lift-out sample(TOMO_001) EMDB-35653: cryo-electron tomogram from mouse islets lift-out sample(TOMO_002) EMDB-35654: cryo-electron tomogram from mouse islets lift-out sample(TOMO_003) EMDB-35655: cryo-electron tomogram from mouse islets lift-out sample(TOMO_004) |
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