PřF:C9940 3-Dimensional Transmission Ele - Course Information
C9940 3-Dimensional Transmission Electron Microscopy (3DEM)Faculty of Science
- Extent and Intensity
- 2/0/2. 2 credit(s) (plus extra credits for completion). Type of Completion: zk (examination).
- Mgr. Jiří Nováček, Ph.D. (lecturer)
Dr. rer. nat. Jürgen Plitzko (lecturer)
Dr. Tanvir Shaikh (lecturer)
- prof. RNDr. Vladimír Sklenář, DrSc.
National Centre for Biomolecular Research - Faculty of Science
Supplier department: National Centre for Biomolecular Research - Faculty of Science
- Course Enrolment Limitations
- The course is offered to students of any study field.
- Course objectives
- 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.
- 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
- Teaching methods
- Lectures, seminars and tutorials
- Assessment methods
- Oral examination
- Language of instruction
- Further Comments
- Study Materials
The course is taught annually.
The course is taught: in blocks.