* Load is given in academic hour (1 academic hour = 45 minutes)
1. Choose the appropriate method for the preparation of biological samples for transmission electron microscopy.
2. Choose the appropriate method for the preparation of biological samples for scanning electron microscopy.
3. Apply and use methods for the preparation of biological samples for research of cells and tissues ultrastructure.
4. Analyze and interpret biological ultrastructures obtained by the above mentioned techniques and assess their quality.
5. Edit and display the results of research in the form of scientific work.
6. Edit and display the results of research in the form of oral communication.
It is the advanced course in which students will get a detailed insight into the basic principles and application of transmission and scanning electron microscopy. Students will acquire basic knowledge on preparation of biological samples for transmission and scanning electron microscopy.
1. The basic principles of electron microscopy (EM);
2. Components of transmission electron microscope (TEM);
3. Electron gun, types of cathodes, electromagnetic lenses and system for vacuum achievement in TEM
4. Types of electron interaction with the sample;
5. Preparation of biological samples for TEM (chemical fixation and fixation by freezing, dehydration, embedding, sectioning);
6. Contrast methods in TEM (positive and negative staining, metal shadowing);
7. Production of replicas and method of fracturing / etching of frozen material;
8. High voltage electron microscopy;
9. Components of scanning electron microscope (SEM);
10. Types of signals in the SEM;
11. Preparation of biological samples for SEM (fixation, dehydration, drying, applying conductive layer);
12. Special types of SEM (Low vacuum SEM, Environmental SEM);
13. Application of electron microscopy in biological research (immunocytochemistry, enzymatic cytochemistry, autoradiography) and image analysis in electron microscopy;
14. Analytical methods in EM (X-ray analysis, electron diffraction, EELS);
15. Scanning Probe Microscopes (atomic force microscopy, scanning tunneling microscopy).
1. Building up the experiment;
2. Fixation, postfixation and dehydration of biological tissues /cells for transmission electron microscope;
3. Use of transitional solvents and embedding biological samples in epoxy resin;
4. Crafting the glass knives;
5. Making support films and sputter coating the grids for electron microscopy;
6. Negative staining and metal shadowing using vacuum: viruses, macromolecules and particles;
7. Sectioning the material with the ultramicrotome;
8. Making and contrasting the semi thick sections;
9. Making and contrasting the ultrathin sections;
10. Work with the transmission electron microscope;
11. Preparation of materials for scanning electron microscope;
12. Work with the scanning electron microscope;
13. Recording and production of the micrographs of various biological samples;
14. Analysis and interpretation of biological structures and ultrastructures;
15. Measurements, morphometry and computerized image analysis.
- Bozzola, J.J., Russell, L.D. (1998): Electron Microscopy. Principles and Techniques for Biologist. 2nd Edition, Jones and Bartlett Publishers.
- Robards, A.W., Wilson, A.J. (1993): Procedures in Electron Microscopy. John Wiley & Sons Ltd.
- Glauert, A.M. (ed.) (1972-1985): Practical methods in electron microscopy. Vol. 1-10, Elsevier, Amsterdam, New York, Oxford.
- Znanstveni članci koji se odnose na istraživanje ultrastrukture stanica i tkiva.
- Plattner, H., Zingsheim, H.P. (1987): Elektronenmikroskopische Metodik in der Zell- und Molekularbiologie. G. Fischer Verlag, Stuttgart, New York.