C9940 3-Dimensional Transmission Electron Microscopy (3DEM)

Přírodovědecká fakulta
jaro 2020
Rozsah
2/0/2. 2 kr. (plus ukončení). Ukončení: zk.
Vyučující
Mgr. Jiří Nováček, Ph.D. (přednášející)
Dr. rer. nat. Jürgen Plitzko (přednášející)
Dr. Tanvir Shaikh (přednášející)
Garance
prof. RNDr. Vladimír Sklenář, DrSc.
Národní centrum pro výzkum biomolekul – Přírodovědecká fakulta
Dodavatelské pracoviště: Národní centrum pro výzkum biomolekul – Přírodovědecká fakulta
Omezení zápisu do předmětu
Předmět je otevřen studentům libovolného oboru.
Cíle předmětu
Transmission electron microscopy (TEM) in its various flavours is nowadays an established characterization tool for structures in life as well as in material science. Especially in todays ‘nano-world’ it is, by and large the only technique for an in-depth visual investigations at the nanometer scale and even in the sub-nanometer regime. The study of complex materials and hybrid compounds, and their interfaces and defects by TEM was and is a major focus in material science. Moreover, three-dimensional (3D) electron microscopy (3DEM) has a particularly important role in the repertoire of techniques strcutural biologists use for probing the structure and functions of molecules and macromolecular complexes in their functional cellular context. 3DEM in life sciences comprises three major branches: cryo-electron crystallography, cryo-electron microscopy of purified single particles (where ‘particles’ stand for proteins and macromolecular complexes) and cryo-electron tomography (the three dimensional investigation of any non periodic - ‘pleiomorphic’ - object). This course will explain the basics of transmission electron microscopy, electron tomography and single particle cryo-EM. It will include the experimental setups and instrumental prerequisites for 3D work, and their actual implementation. Specimen preparation methods suited for biological applications will be described and explained in detail. Since all tomographic methods are based on different reconstruction algorithms, and elaborate image processing and visualization routines, they will be included in this tutorial as well. At the end of the course students should understand the principles of structural analysis by TEM and its application in chemistry, biochemistry, structural biology, biophysics and materials science.
Osnova
  • 1. Introduction: TEM history and contemporary trends - applications of TEM to structural studies. 2. Introduction to electron optics, image formation, diffraction and automation. 3. Specimen preparation, negative stain, thin-film vitrification, high-pressure freezing, cryo-sectioning and modern micromachining methods. 4. Radiation effects; Introduction to image analysis; Sources of noise. 5. Principle and background of Fourier analysis, contrast transfer function (CTF), convolution and cross-correlation. 6. Electron tomography. 7. Principles of image analysis and 3D reconstruction methods 8. Single Particle analysis and image processing 9. 3D Visualization methods 10. Hybrid Methods: Combining X-ray crystallography and cryo-EM, combining light-microscopy and cryo-EM
Výukové metody
Lectures, seminars and tutorials
Metody hodnocení
Oral examination
Vyučovací jazyk
Angličtina
Další komentáře
Předmět je vyučován každoročně.
Výuka probíhá blokově.
Předmět je zařazen také v obdobích jaro 2012, jaro 2012 - akreditace, jaro 2013, jaro 2014, jaro 2015, jaro 2016, jaro 2017, jaro 2018, jaro 2019, jaro 2021, jaro 2022, jaro 2023, jaro 2024, jaro 2025.