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- EMDB-35655: cryo-electron tomogram from mouse islets lift-out sample(TOMO_004) -
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Open data
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Basic information
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Title | cryo-electron tomogram from mouse islets lift-out sample(TOMO_004) | |||||||||
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![]() | islet / vesicle / insulin / EXOCYTOSIS | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | electron tomography / cryo EM | |||||||||
![]() | Wu Y / Qin C / Du W / Chen L / Guo Q | |||||||||
Funding support | 1 items
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![]() | ![]() Title: A practical multicellular sample preparation pipeline broadens the application of in situ cryo-electron tomography. Authors: Yichun Wu / Changdong Qin / Wenjing Du / Zhenxi Guo / Liangyi Chen / Qiang Guo / ![]() 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. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 770.1 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 8.9 KB 8.9 KB | Display Display | ![]() |
Images | ![]() | 161.7 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
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Voxel size | X=Y=Z: 15.84 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
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Sample components
-Entire : mouse islets lift-out sample
Entire | Name: mouse islets lift-out sample |
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Components |
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-Supramolecule #1: mouse islets lift-out sample
Supramolecule | Name: mouse islets lift-out sample / type: tissue / ID: 1 / Parent: 0 |
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Source (natural) | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | electron tomography |
Aggregation state | tissue |
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Sample preparation
Buffer | pH: 7.4 |
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Vitrification | Cryogen name: NITROGEN / Details: high pressure freezing. |
High pressure freezing | Instrument: OTHER Details: The value given for _em_high_pressure_freezing.instrument is Leica EM HP100. This is not in a list of allowed values {'BAL-TEC HPM 010', 'LEICA EM HPM100', 'LEICA EM PACT', 'LEICA EM PACT2', ...Details: The value given for _em_high_pressure_freezing.instrument is Leica EM HP100. This is not in a list of allowed values {'BAL-TEC HPM 010', 'LEICA EM HPM100', 'LEICA EM PACT', 'LEICA EM PACT2', 'OTHER', 'EMS-002 RAPID IMMERSION FREEZER'} so OTHER is written into the XML file. |
Sectioning | Focused ion beam - Instrument: OTHER / Focused ion beam - Ion: OTHER / Focused ion beam - Voltage: 30 / Focused ion beam - Current: 0.03 / Focused ion beam - Duration: 1800 / Focused ion beam - Temperature: 81 K / Focused ion beam - Initial thickness: 10000 / Focused ion beam - Final thickness: 310 Focused ion beam - Details: The value given for _em_focused_ion_beam.instrument is Aquilos. This is not in a list of allowed values {'DB235', 'OTHER'} so OTHER is written into the XML file. |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 2.24 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 7.0 µm / Nominal defocus min: 5.0 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Final reconstruction | Number images used: 49 |
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