The Imaging Centre at EMBL Heidelberg gives access to state-of-the-art cryo-EM equipment for structure determination projects using the latest technology and methods in single-particle analysis, cryo-electron tomography (cryo-ET) and Cryo-correlative light and electron microscopy (Cryo-CLEM).
After cryofixation by plunge or high pressure freezing, samples can be imaged in a cryo-confocal microscope (Zeiss LSM 900 Airyscan2) and subsequently processed further in a cryo-FIB-SEM (Zeiss Crossbeam 550) using a common sample mount and cryo-transfer system (Quorum) that maintains sample orientation between the LM and EM systems so that coordinates of target structures are efficiently transferred. FIB-milling of target areas allows the creation of lamellae that can be subsequently imaged by tomography in a 300 kV Titan Krios G4 TEM.
Cryo-grids will be screened with a Thermo Fisher Scientific Glacios equipped with a Falcon 4 electron counting direct electron detector. High-end data acquisition will be performed with a Thermo Fisher Scientific Titan Krios with a three-condenser lens system. It features the latest technology for high-end single-particle acquisition including a cold field emission gun (C-FEG), SelectrisX energy filter, Falcon 4 electron counting camera and volta phase plate. Both microscopes are equipped with an automatic sample loading system and configured for high-throughput automatic data acquisition.
Cryo-grids will be screened with a Thermo Fisher Scientific Glacios equipped with a Falcon 4 electron counting direct electron detector. High-end data acquisition will be performed with a Thermo Fisher Scientific Titan Krios with a three-condenser lens system. It features the latest technology for tomography data acquisition including a cold field emission gun (C-FEG), SelectrisX energy filter, Falcon 4 electron counting camera and volta phase plate. Both microscopes are equipped with an automatic sample loading system and configured for high-throughput automatic data acquisition.
The EMBL Heidelberg operates the Imaging Centre, a user facility providing cryo-EM services. EM engineers and application specialists will help and support any scientist with cryo-EM data collection to tackle a broad range of biological questions and projects based on single-particle analysis and cryo-electron tomography for high-resolution structural determination of macromolecular complexes. We also provide support on direct correlation between confocal fluorescence imaging and cellular tomography under cryo conditions (cryo-CLEM). The centre is also offering remote service to all interested users.
Currently we offer access to the following services:
1. High-end data collection for cryo-EM single-particle analysis
2. High-end data collection for cryo-electron tomography
3. Cryo-confocal imaging and cellular tomography under cryo conditions
Specific equipment:
Thermo Fisher Scientific Glacios equipped with a Falcon 4 electron counting direct electron detector;
Thermo Fisher Scientific Titan Krios with a Falcon 4 electron counting camera and volta phase plate;
LSM900 Airyscan2 equipped with an Linkam cryo stage and configured for high-resolution confocal imaging of fluorescent sample;
ZEISS Crossbeam 550 equipped to generate cryo-lamellae;
The centre and its instruments are available for the entire scientific Instruct-ERIC community.
For more details on instruments, methods, experts, how to apply, please visit our website.
