Technology Network HDcryoNet
The Cryo-Electron Microscopy Network of Heidelberg University
The Cryo-Electron Microscopy Network of Heidelberg University (HDcryoNet) hosts cryo-EM infrastructure located at the BioQuant and at the Biochemistry Center. The aim of HDcryoNet is to foster collaborative research relying on cryo-EM methods, and to teach these methods to students at Heidelberg University. HDcryoNet operates as a network of users who independently use the equipment and share duties. HDcryoNet operates four high-end cryo-EM instruments.
Instruments located at the BioQuant
Aquilos cryo-FIB/SEM
The Aquilos dual-beam microscope is a dedicated instrument for the preparation of thin cryo-lamellae of vitrified cells that are subsequently studied by cryo-electron tomography in the Krios cryo-TEM. The microscope is equipped with a focused Gallium ion beam column, an electron beam column, and a cryo-stage. It uses an Edward-Thornley detector and in lens and in column Trinity detection systems for detection of secondary and backscattered electrons. The microscope can be operated in a semi-automated fashion using Autoscript scripting. The microscope is located in the BSL2 suite together with the Krios microscope to support in situ cryo-electron tomography projects. The microscope was acquired through DFG funding and is in operation since 2018.
Building: INF 229
Room: U42&45 (Biosafety level 2)
Phone: +49 6221 5419373
Krios – cryo-TEM (300kV)
The Krios cryo-transmission electron microscope is usually operated at 300kV and is dedicated to cryo-electron tomography and high-resolution data collection for single-particle analysis. It is equipped with a field emission gun, K3 Direct Electron Detector, Quanta GIF imaging filter, Volta phase plate and a dual-axis stage. Recently it has been upgraded with a state-of-the-art 3rd generation Autoloader. Data collection is carried out remotely using EPU or SerialEM. The microscope is located in the BSL2 suite together with the Aquilos microscope. The microscope was acquired through CellNetworks funding and is in operation since 2012.
Building: INF 229
Room: U41 (Biosafety Level 2)
Phone: +49 6221 5419370
Instruments located at BZH
Glacios – cryo-TEM (200kV)
The Glacios is a state-of-the-art 200kV cryo-transmission electron microscope equipped with an Autoloader that allows streamlined sample screening and data collection for single-particle analysis of purified complexes. It is equipped with a field emission gun, Falcon3-EC direct detector, and a Volta phase plate. Data collection is carried out remotely using EPU or SerialEM. The microscope is located in vicinity of the Talos L120C microscope to support single-particle projects. The microscope was acquired through DFG funding and is in operation since 2020.
Building: INF 328
Phone: +49 6221 544377
Talos L120C – (cryo)-TEM 120 kV
Talos L120C is a state-of-the-art 120kV transmission electron microscope that can be operated at cryogenic temperatures. It is equipped with a LaB6 electron source and a Ceta-M camera with a 4k × 4k CMOS sensor. This highly versatile microscope is dedicated to sample screening at both room and cryogenic temperatures. It operates with side-entry holders (several holders are available, incl. the newest Elsa Cryo-Transfer Holder). The microscope was acquired through DFG funding and is in operation since 2019.
Building: INF 328
Phone: +49 6221 544165
Cryo-EM sample preparation instruments
Automatic plunge freezers:
Vitrobot Mark IV (BioQuant), Vitrobot Mark IV (BZH), Gatan Cryoplunge 3 (BioQuant), Leica GP2 (CIID)
Glow dischargers:
Gatan Solarus 950 (BioQuant)
Data storage and processing infrastructure
The computing center of Heidelberg University provides access to a fast large scale data storage facility (Scientific Data Storage) and a high-performance computing cluster (bwForCluster) equipped with multi-GPU nodes tailored for cryo-EM data analysis.
More information on the high-performance computing cluster (bwForCluster):
Further information and training related to computational resources available on campus at Heidelberg University are provided by the Community Forum Data-Intensive Computing and the HPC-Support.
CONTACT
For inquiries, please contact hdcryonet@uni-heidelberg.de
STEERING COMMITTEE
Current members of the HDcryoNet steering committee (ordered alphabetically):
- Petr Chlanda (BioQuant/Department of Infectious Diseases-Virology)
- Cristina Paulino (BZH)
- Stefan Pfeffer (ZMBH)
- Rasmus R. Schröder (BioQuant)
- Irmi Sinning (BZH)
- User Group
user group representatives
- Dirk Flemming (BZH)
- Götz Hofhaus (BioQuant)
- Jan Rheinberger (BZH)
CRYO-EM Gallery
Recent Publications
Vermeulen BJ, Böhler A, Gao Q, Neuner A, Župa E, Chu Z, Würtz M, Jäkle U, Gruss OJ, Pfeffer S, Schiebel E. γ-TuRC asymmetry induces local protofilament mismatch at the RanGTP-stimulated microtubule minus end. EMBO J. 2024 May;43(10):2062-2085. doi: 10.1038/s44318-024-00087-4. Epub 2024 Apr 10. PMID: 38600243; PMCID: PMC11099078.
Bohl V, Hollmann NM, Melzer T, Katikaridis P, Meins L, Simon B, Flemming D, Sinning I, Hennig J, Mogk A. The Listeria monocytogenes persistence factor ClpL is a potent stand-alone disaggregase. Elife. 2024 Apr 10;12:RP92746. doi: 10.7554/eLife.92746. PMID: 38598269; PMCID: PMC11006417.
Uckeley ZM, Duboeuf M, Gu Y, Erny A, Mazelier M, Lüchtenborg C, Winter SL, Schad P, Mathieu C, Koch J, Boulant S, Chlanda P, Maisse C, Brügger B, Lozach PY. Glucosylceramide in bunyavirus particles is essential for virus binding to host cells. Cell Mol Life Sci. 2024 Feb 1;81(1):71. doi: 10.1007/s00018-023-05103-0. PMID: 38300320; PMCID: PMC10834583.
Winter SL, Chlanda P. The Art of Viral Membrane Fusion and Penetration. Subcell Biochem. 2023;106:113-152. doi: 10.1007/978-3-031-40086-5_4. PMID: 38159225.
Theuerkauf SA, Herrera-Carrillo E, John F, Zinser LJ, Molina MA, Riechert V, Thalheimer FB, Börner K, Grimm D, Chlanda P, Berkhout B, Buchholz CJ. AAV vectors displaying bispecific DARPins enable dual-control targeted gene delivery. Biomaterials. 2023 Dec;303:122399. doi: 10.1016/j.biomaterials.2023.122399. Epub 2023 Nov 16. PMID: 37992599; PMCID: PMC10721713.
Zimmermann L, Zhao X, Makroczyova J, Wachsmuth-Melm M, Prasad V, Hensel Z, Bartenschlager R, Chlanda P. SARS-CoV-2 nsp3 and nsp4 are minimal constituents of a pore spanning replication organelle. Nat Commun. 2023 Nov 30;14(1):7894. doi: 10.1038/s41467-023-43666-5. PMID: 38036567; PMCID: PMC10689437.
Schmidt N, Ganskih S, Wei Y, Gabel A, Zielinski S, Keshishian H, Lareau CA, Zimmermann L, Makroczyova J, Pearce C, Krey K, Hennig T, Stegmaier S, Moyon L, Horlacher M, Werner S, Aydin J, Olguin-Nava M, Potabattula R, Kibe A, Dölken L, Smyth RP, Caliskan N, Marsico A, Krempl C, Bodem J, Pichlmair A, Carr SA, Chlanda P, Erhard F, Munschauer M. SND1 binds SARS-CoV-2 negative-sense RNA and promotes viral RNA synthesis through NSP9. Cell. 2023 Oct 26;186(22):4834-4850.e23. doi: 10.1016/j.cell.2023.09.002. Epub 2023 Oct 3. PMID: 37794589; PMCID: PMC10617981.
Castillo Duque de Estrada NM, Thoms M, Flemming D, Hammaren HM, Buschauer R, Ameismeier M, Baßler J, Beck M, Beckmann R, Hurt E. Structure of nascent 5S RNPs at the crossroad between ribosome assembly and MDM2-p53 pathways. Nat Struct Mol Biol. 2023 Aug;30(8):1119-1131. doi: 10.1038/s41594-023-01006-7. Epub 2023 Jun 8. PMID: 37291423; PMCID: PMC10442235.
Castillo Duque de Estrada NM, Thoms M, Flemming D, Hammaren HM, Buschauer R, Ameismeier M, Baßler J, Beck M, Beckmann R, Hurt E. Structure of nascent 5S RNPs at the crossroad between ribosome assembly and MDM2-p53 pathways. Nat Struct Mol Biol. 2023 Aug;30(8):1119-1131. doi: 10.1038/s41594-023-01006-7. Epub 2023 Jun 8. PMID: 37291423; PMCID: PMC10442235.
For a comprehensive overview of publications, please see document below.