* Load is given in academic hour (1 academic hour = 45 minutes)
1. Overmaster the basic skills necessary for independent elaboration and interpretation of light microscopy samples.
2. Choose the appropriate method for the preparation of biological samples for transmission electron microscopy.
3. Choose the appropriate method for the preparation of biological samples for scanning electron microscopy.
4. Apply and use methods for the preparation of biological samples for research cells and tissues ultrastructure.
5. Analyze and interpret biological ultrastructures obtained by the above mentioned techniques and assess their quality.
It is the introductory course in which students will gain insight into the basic principles and application of special light microscopy, transmission and scanning electron microscopy. Students will acquire basic knowledge on preparation of biological samples for transmission and scanning electron microscopy.
1. Physical basis of light microscopy;
2. Components of light microscope;
3. The basic principles and application of specific types of light microscopes (dark field, phase contrast, differential interference contrast, polarization microscopy, fluorescence microscopy, confocal laser scanning microscopy);
4. The basic principles and application of specific types of light microscopes (polarization microscopy, fluorescence microscopy, confocal laser scanning microscopy);
5. Physical basis of electron microscopy (EM);
6. The basic principles of transmission electron microscopy (TEM);
7. The basic components and working principle of TEM;
8. Basic methods of preparation of biological samples for TEM;
9. Special methods of preparation of biological samples for TEM;
10. The basic principles of scanning electron microscopy (SEM);
11. The basic components and working principle of SEM;
12. Preparation of biological samples for SEM;
13. Fundamentals of other types of electron microscopy (high voltage EM, Environmental SEM, Low vacuum SEM);
14. Analytical methods in EM (X-ray analysis, electron diffraction, EELS);
15. Scanning Probe Microscopes (atomic force microscopy, scanning tunneling microscopy).
1. Components of the light microscope, resolving power of the microscope and the microscope as a measuring instrument;
2. Application of phase contrast microscope and differential interference contrast microscope;
3. Application of polarizing microscope and dark filed microscope;
4. Application of fluorescence microscope and confocal laser scanning microscope;
5. Fixation, postfixation and dehydration of biological tissues/cells for transmission electron microscope;
6. Use of transitional solvents and embedding biological samples in epoxy resin;
7. Crafting the glass knives;
8. Making support films and sputter coating the grids for electron microscopy;
9. Negative staining and metal shadowing using vacuum: viruses, macromolecules and particles;
10. Sectioning the material with the ultramicrotome. Making and contrasting the semi thick and ultrathin sections;
11. Work with the transmission 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 ultrastructures;
15. Measurements, morphometry and computerized image analysis.
- Abramowitz M. (2003): Microscope. Basics and Beyond. (http://micro.magnet.fsu.edu/primer/pdfs/basicsandbeyond.pdf)
- Bredbury, S. (1989): Introduction to the Optical Microscopy, Revised Edition, Oxford Univ. Press.
- 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.
- 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.