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Cryo-EM Platform (冷冻电镜平台 )

Cryo-EM technology is an important developmental step for the National Protein Science Research (Beijing) Facility based at Tsinghua. Cryo-EM platform mainly uses Cryo-EM 3D reconstruction technology to make 3D structural analysis of cells, subcellular organelles, biological macromolecules and complexes at several levels, to find answers to questions arising in bioscience, thus organically integrating cell biology and structural biology. TEM and its variants are the main equipment platforms used for operations at the cellular level. These are suitable for single particle analysis, electron tomography and electron crystallography research. Furthermore, the platform has a complete set of sample preparation systems including quick freezer, high vacuum turbo evaporator and plasma cleaner used for ultrastructural study.

 

 For detailed information and charges, please visit Protein Research and Technology Center's Protein Cryo-EM Platform website: http://phoenix.tsinghua.edu.cn/platform?group=17. 

 Contact us: Room 125, Life Science Building, Tsinghua University, Haidian District, Beijing.

 Contact person:

Jianlin Lei (General Director)jllei@biomed.tsinghua.edu.cn62797699
Lingpeng Cheng (Technical Director)
lingpengcheng@tsinghua.edu.cn
62773238
Yanji Xuxuyanji@tsinghua.edu.cn
62772234
Xiaomei Lilixiaomei@biomed.tsinghua.edu.cn
62772234
Mingfang Wang wangmingfang@biomed.tsinghua.edu.cn62772234
Xiaomin Lixiaominli@tsinghua.edu.cn
62772326
Bin Xu 
binxu@biomed.tsinghua.edu.cn
62772326


FEI Titan Krios 300KV FETEM (D3172)  

Model
FEI Titan Krios 300KV
Manufacturer
FEI
1.png
Filament type
ZrO/W (100) Schottky field emission gun (S-FEG)
Objective lens type
C-Twin
Point resolution
0.25nm
Information resolution limit
0.14nm
Accelerating voltage
80kV –300kV
Magnification range
40´ – 740k´ 
Maximum tilt angle of samples
±71.5°, samples can rotate 90 degrees in horizontal plane
Digital camera

Gatan K2 Summit direct electron detector

Gatan Orius SC600B (831) CCD

FEI Eagle 4k CCD

Other characteristics

Autoloader system can at the most load and store 12 samples at one time

Three-level condenser laminating system ensures wide-range of adjustable parallel lighting 

Constant power mode for magnetic lens system guarantees high imaging stability

No image rotation under different magnification

Automatic liquid nitrogen filling

Feasible for remote operations

Phase Plate can greatly improve image contrast

HAADF (high-angle annular dark field) used in STEM mode

Integrated iCorr Interactive System

• Storage Location: Room 125, 129, Life Science Building


FEI Titan Krios 300KV Field emission scanning energy filtering STEM (D3418)

ModelFEI Titan Krios 300KV 
Manufacturer

FEI

2.png
Filament typeZrO/W (100) Schottky field emission gun (S-FEG)
Objective lens typeC-Twin
Point resolution0.25nm
Information resolution limit0.14nm
Accelerating voltage80kV –300kV
Magnification range40´ – 740k´
Maximum tilt angle of samples±71.5°, samples can rotate 90 degrees in horizontal plane
Digital camera

FEI Falcon II direct electron detector

Gatan Orius SC200B (830) CCD 

Energy filtering systemGIF Quantum 964
Other characteristics

Autoloader system can at the most load and store 12 samples at one time

Three-level condenser laminating system ensures wide-range of adjustable parallel lighting

Constant power mode for magnetic lens system guarantees high imaging stability

No image rotation under different magnification

Automatic liquid nitrogen filling

Feasible for remote operations

HAADF (high-angle annular dark field) used in STEM mode

• Storage Location: Life Science Building yard


FEI Titan Krios 300KV Cs-correct field emission scanning TEM (D3424)

ModelFEI Titan Krios 300KV3.png
ManufacturerFEI
Filament typeSchottky field emission gun (X-FEG)
Objective lens typeC-Twin
Information resolution limit0.14nm
Accelerating voltage80kV –300kV
Magnification range40´ – 740k´
Maximum tilt angle of samples±71.5°, samples can rotate 90 degrees in horizontal plane
Digital camera

FEI Falcon II direct electron detector

Gatan Orius SC200B (830) CCD 

Cs-correct systemImage Cs-corrector
Other characteristics

Autoloader system can at the most load and store 12 samples at one time

Three-level condenser laminating system ensures wide-range of adjustable parallel lighting

Constant power mode for magnetic lens system guarantees high imaging stability

No image rotation under different magnification

Automatic liquid nitrogen filling

Feasible for remote operations

Phase Plate can greatly improve image contrast

• Storage Location: Life Science Building yard


FEI Tecnai Arctica 200KV FETEM (D683)

ModelFEI Tecnai Arctica
ManufacturerFEI4.png
Filament typeZrO/W (100) Schottky field emission gun (S-FEG)
Objective lens typeC-Twin
Point resolution0.3nm
Information resolution limit0.23nm
Accelerating voltage20kV –200kV
Magnification range21.5´ – 560k´
Maximum tilt angle of samples±71.2°
Digital camera

FEI Falcon II direct electron detector

Gatan Orius SC200B (830) CCD

Other characteristics

Autoloader system can at the most load and store 12 samples at one time

Automatic liquid nitrogen filling

Feasible for remote operations

Phase Plate can greatly improve image contrast

• Storage Location: Room 119, 121, Life Science Building


FEI Tecnai F20 200KV FESEM (D545)

ModelFEI Tecnai F20 Twin5.png
ManufacturerFEI
Filament typeZrO/W (100) Schottky field emission gun (S-FEG)
Objective lens typeTwin
Point resolution0.27nm
Line resolution0.144nm
Information resolution limit0.16nm
Accelerating voltage20kV –200kV
Magnification range21´ – 700k´
Maximum tilt angle of samples±80°
Available sample holder

Normal-temperature common monoclinal holder

Normal-temperature Tomography sample holder, normal-temperature multi-sample holder

Gatan 626 60° freezing sample holder, Gatan 626 70°freezing sample holder

Gatan CT3500 freezing sample holder, Gatan 914 Tomography freezing sample holder

Digital camera

DE-20 direct electron detector (Direct Electron, LP)

Gatan US4000 (895) CCD、Gatan Orius SC200B (830) CCD

Other characteristicsHAADF (high-angle annular dark field) used in STEM mode

• Storage Location: Room 126, Life Science Building 


FEI Tecnai Spirit BioTwin 120KV TEM (D1266)

ModelFEI Tecnai Spirit BioTwin
ManufacturerFEI6.png
Filament typeW or LaB6
Objective lens typeBioTwin
Point resolution0.49nm
Line resolution0.344nm
Accelerating voltage20kV –120kV
Magnification range22´ – 340k´
Maximum tilt angle of samples±80°
Available sample holder

Normal-temperature common monoclinal holder

Normal-temperature Tomography sample holder

Normal-temperature multi-sample holder

Gatan 626 60° freezing sample holder, Gatan 626 70°freezing sample holder

Gatan CT3500 freezing sample holder, Gatan 914 Tomography freezing sample holder

Digital cameraGatan US4000 (895) CCD

• Storage Location: Room 124, Life Science Building


FEI Tecnai Spirit Twin 120KV TEM (D1319)

ModelFEI Tecnai Spirit Twin
ManufacturerFEI7.png
Filament typeW or LaB6
Objective lens typeTwin
Point resolution0.344nm
Line resolution0.204nm
Accelerating voltage20kV –120kV
Magnification range21´ – 740k´
Maximum tilt angle of samples

-80°,90.5°


Available sample holder

Normal-temperature common monoclinal holder

Normal-temperature Tomography sample holder

Normal-temperature multi-sample holder

Gatan 626 60° freezing sample holder, Gatan 626 70°freezing sample holder

Gatan CT3500 freezing sample holder, Gatan 914 Tomography freezing sample holder

Digital cameraFEI Eagle 4k CCD
Other characteristicsIntegrated iCorr Interactive System

• Storage Location: Room 121, Life Science Building


The platform also has a Biological EM Sample Preparation system, including:


Quick freezing instrument: Quickly freezing solution samples on micro grids for Cryo-EM

Model: FEI Vitrobot Mark IV,Gatan CP3,Leica EM GP

Plasma cleaner: hydrophilic micro grids and clean sample holders

Model: Harrick PDC-32G-2 Basic,Gatan Solarus

High vacuum coater: Used to apply carbon coating and various metal films

Model: Cressington 208C,Denton Vacuum DV-502


Cell Cryo-EM Platform

The combination of EM sample preparation and EM technology is the commonest technology used in cellular studies and tissue ultrastructure. It can be applied to IEM research and for electron tomography 3D imaging. Quality of cell samples in EM is a critical factor affecting quality of EM images. This platform currently has 5 cell EM sample preparation devices, including 3 Ultramicrotomes, 1 high-pressure freezer, and 1 freezing replacer. One FEI Tecnai Spirit BioTwin120KV TEM is included for ultrathin section observation. EM cell sample preparation technical services provided include:

(1) Cryosection preparation (Tokuyaso method/ CEMOVIS method); (2) Immunolabeling; (3) High-pressure high-speed cryofixation; (4) low-temperature dehydrated embedding; (5) CLEM; (6) Ultrathin section observation and 3D reconstruction analysis.


For detailed information and charges, please visit the Protein Research and Technology Center’s Cell EM Platform website: http://phoenix.tsinghua.edu.cn/platform?group=64.

 

High-pressure Freezer

A high-pressure freezer can do sample cryofixation under high pressure. It is dedicated auxiliary EM sample preparation equipment. Compared to chemical fixation, cryofixation can be completed in milliseconds without resolution loss, and capture every moment of physiological activity at the cellular level, thus maximally preserving initial structure and viability of protein, enzyme and antigen. It also retains intrinsic permeability of membrane system, and fixes soluble micromolecules and ions. This equipment can be applied to cell biology, neurobiology, immunology, stem cell biology and other allied research fields.

Application scope: Cryofixation of samples with high water content, also samples that are difficult to be fixed using chemicals; (2) CLEM; (3) CEMOVIS sample preparation.

ModelLeica EM HPM1008.png
ManufacturerLeica
Effective cryofixation thicknessAt most 200μm
Pressure imposed in cryofixationAbout 2100 bar
Freezing rate12000K/s~25000K/s
Sample holderIncluding 3mm/6mm CLEM, freeze-fracture system, pipe system (Tokuyaso/CEMOVIS sample preparation), membrane system


Freeze Substitution

Freeze Substitution equipment is required for preparing samples after high-pressure freezing. It can complete dehydration substitution at low temperature, and allows low temperature embedding and resin polymerization. After sample freeze substitution, temperature can be restored to normal for normal-temperature embedding. Application scope: (1) Freeze substitution technology; (2) PLT technology; (3) Low temperature embedding technology.

ModelLeica EM AFS29.png
ManufacturerLeica
Temperature scopeControllable under -140°C ~ +70°C
UV polymerizationBuilt-in UV LED lamp can automatically complete UV polymerization
Sample embeddingNormal/ low temperature


Ultramicrotome

It is dedicated auxiliary sample preparation equipment used for preparing biomedical (Tokuyaso/CEMOVIS methods), high polymer ultra-thin sample sections.

Application scope: Prepare semi-thin sections, ultra-thin sections, ET and 3D reconstruction sections at normal/low temperature.

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ModelUltracut R      EM UC7+FC7      EM UC7+FC7+ micromanipulator
ManufacturerLeica
Section thickness1nm~15μm
Working temperatureNormal microtome: room temperature; freezing microtome: -185°C ~ -15°C

 

TEM

FEI Tecnai G2 Spirit BioTwin TEM is a completely digitally controlled TEM, with perfect EM stability, repeatability, simple to operate and convenient management. Equipped with a BioTwin type objective lens the EM features high imaging contrast and can realize high-level of automation. It is suitable for single particle analysis, electron tomography and electron crystallography research.

Application scope: (1) Biomacromolecule negatively stained and frozen sample checking and data collection; (2) Ultrathin section sample checking and data collection; (3) Ultrastructure tomography analysis of biological samples.

ModelFEI Tecnai Spirit BioTwin11.png
ManufacturerFEI
Filament typeW or LaB6
Point resolution0.49nm
Line resolution0.344nm
Accelerating voltage20kV – 120kV
Magnification range22´ – 340k´
Maximum tilt angle of samples±80°
Digital camera

FEI Eagle 4k CCD, 

Gatan Orius SC1000 (832) CCD

• Storage Location: TEM--Room E101, Zhengyutong Mecidal Building; Others--Room E220, Zhengyutong Mecidal Building

• Contact Person: Ying Li

• Tel: +86-10-62771601

• Email: liying@biomed.tsinghua.edu.cn 

中心地址:北京市海淀区清华大学