Cryo-electron Microscopy and Tomography

Modern electron microscopy in structural biology on the cellular and molecular level is performed by cryo-electron microscopy and cryo-electron tomography. Cryo-electron tomography (cryo-ET) is the only method to address pleiomorphic structures like cells and organelles in a close to native state, while cryo-electron microscopy (cryo-EM) is applied to study single particles, primarily larger macromolecular complexes, which have been isolated and purified by biochemical methods. Both methodologies provide information on the cellular and molecular level and are therefore ideal for in-depth structural-functional analysis in combination with state of the art biochemical characterisation. The main activities of the Core Facility will be centred on application of cryo-EM and cryo-ET, implementations of the required image processing capabilities, and explorations of suitable cryo-preparation techniques.

Key Equipment

  • High-end electron microscope for high-resolution cryoEM and cryo-tomography FEI Titan Krios (80 - 300 kV) equipped with an energy filter and a direct detector camera
  • Conventional cryoEM microscope FEI F20 (200 kV) equipped with a CCD camera and a dual beam FIB/SEM instrument (Versa3D) for micromachining of thin lamellas of vitrified cells usable for electron tomography
  • Equipment for sample preparation
  • Vitrification robot Vitrobot Mark IV

Expertise

  • Studies, at the highest currently possible level, of cellular structures and organelles, protein complexes and various phenomena at the macromolecular level by 3D imaging

Open-access

  • Advanced and routine application of cellular electron tomography
  • Application of correlative methods integrating light/fluorescence microscopy with high resolution cryo-electron microscopy techniques
  • Single particle analysis
  • Preliminary negative stain/RT- and advanced cryo-electron microscopy methods

Core Facility contact

Jiří Nováček - jiri.novacek@ceitec.muni.cz

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More information about Cryo-electron Microscopy and Tomography and related services you can find in document Equipement and services of CIISB (page 17).


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