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MPI für Biochemie  

Molecular Structural Biology
Wolfgang Baumeister

Instrumentation

 

Members

Dr. Jürgen M. Plitzko

Phone +49 - 89 - 8578 2645

 


Current equipment

The current equipment comprises several dedicated cryo-transmission electron microscopes (see spreadsheet) which are used either for cellular cryo-electron tomography, single particle investigations or both. All microscopes are manufactured by FEI Company, Eindhoven, The Netherlands (former Philips) and all are equipped with digital cameras (CCDs) of different sizes (respectively detection area and pixelsize). All microsopes operate at liquid nitrogen temperature (90K) and the Polara instruments can be even cooled down to liquid helium temperature (10K).

 

Name of instrument Vintage Operation voltage [kV] Energy filter CCD Applications
CM 120 Biofilter 1993 120 GIF1100 Gatan 1024x1024 (24 µm) postGIF Cellular tomography
CM 200 FEG 1991 160/200   TVIPS 4096x4096 (15 µm) Single particle
CM 300 FEG 1998 300 GIF 2000 Gatan 2048x2048 (30 µm) Cellular tomography
Tecnai F20 2007 120/160/200   FEI Eagle 4096x4096 (15 µm) Single particle
Tecnai 'Polara' G2 F30 (1) 2003 300 HR-GIF Gatan 4096x4096 (15 µm) preGIF
Gatan 2048x2048 (30 µm) postGIF		 Cellular tomography
Single particle
Tecnai 'Polara' G2 F30 (2) 2008 300 GIF 2000 Gatan 2048x2048 (30 µm) postGIF Cellular tomography
Titan KRIOS 2007 80-300   TVIPS 8192x8192 (15 µm) Single particle

1 GIF = Gatan Imaging Filter

 

microscopes_ed2

Tecnai F30 Helium (‘Polara’) microscope (left) as installed at the MPI of Biochemistry in Martinsried. This cryo-EM comprises a manual operated multispecimen stage (to load up to 6 samples) and it can be used either at liquid nitrogen or helium temperature. Titan KRIOS, a microscope in a box (middle, right image): the latest generation of computer controlled TEMs dedicated for cryo-EM, combining the latest technology in automation and robotics (especially for sample mounting; here up to 12 samples can be transferred at once in a robot called Autoloader).

In-house developments

The in-house developments include the design and manufacturing of hardware components and the design and implementation of suitable software routines to facilitate and further improve the applications in cryo-EM and especially cryo-ET. Hardware tools encompass the cryo-preparation/cryo-transfer/cryo-storage, dual-axis side entry cryo-holders and accessories for light/fluorescence microscopy at cryo-temperatures (see e.g. Correlative microscopy&FIB). Moreover we are actively involved in the exploration/testing and implementation of new approaches in electron microscopy, where we collaborate with the microscope and CCD camera suppliers. Software tools include the automation for the acquisition and analysis in cryo-EM and cryo-ET (see e.g. Software development).

Publications

  1. Plitzko JM: Cellular cryo-electron tomography (CET)-Towards a voyage to the inner space of cells. In Single Cell Analysis. Edited by Anselmetti D: Wiley-VCH Verlag; 2009:39-68.
  2. Nickell, S, Beck, F, Scheres, SHW, Korinek, A, Forster, F, Lasker, K, Mihalache, O, Sun, N, Nagy, I, Sali, A, Plitzko JM, Carazo, JM, Mann, M, Baumeister, W: Insights into the molecular architecture of the 26S proteasome. Proceedings of the National Academy of Sciences of the United States of America 2009, 109:11943-11947.
  3. Ziegler A, Rockel B, Hegerl R, Freitag B, Lucken U, Plitzko JM: Aberration-corrected microscopy for structural biology applications. Journal of Microscopy-Oxford 2009, 233:170-177.
  4. Plitzko JM, Baumeister W: Cryo-Electron Tomography (CET). In Science of Microscopy. Edited by Hawkes P, Spence J: Springer; 2006:535-604. vol 1.