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Lecturer(s)
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Řehák Jan, Mgr.
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Luptovčiak Ivan, Mgr. Ph.D.
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Ovečka Miroslav, prof. Mgr. Ph.D.
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Hlaváčková Kateřina, Mgr. Ph.D.
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Course content
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1. Principle of light microscopy. Properties of light and function of optical lenses. Construction and operation of optical elements of a light microscope, principle of imaging. Classification and basic types of light microscopes. Exercise 1: Introduction to light microscopy. 2. Optical abberaions, contrast and resolution. Parameters of light microscopy usable in cell research. Principle of setting up Köhler illumination. Exercise 2: Comparison of light microscope and stereomicroscope. 3. Principle of construction of microscope objectives, theory of image formation and magnification. Types of objectives and possibilities of their application in various methods of imaging plant objects. Exercise 3: Making manual sections from plant material. 4. Special techniques of light microscopy I. Contrast in a light microscope, phase, amplitude. Imaging in the dark field, principle of phase contrast and polarization. Exercise 4: Staining of microscopic preparations from plant material. 5. Special techniques of light microscopy II. Principle of differential interference contrast, polarization and other methods of contrast modulation in light microscopy. Exercise 5: Anatomy of the stem and root of monocots and dicots. 6. Reflection, refraction and diffraction of light, interference of light waves, point stread function. Resolution of the microscope. Exercise 6: Storage substances in plant cells. 7. Principle of fluorescence microscopy. Construction of a fluorescence microscope, theory of fluorescence filters, use of fluorochromes. Exercise 7: Preparation of the epidermis of a leaf for microscopy. 8. Application of fluorescence microscopy in the study of plant cells. Autofluorescence, secondary fluorescence, immunofluorescence, use of fluorochromes for in vivo microscopy. Methods of studying plant cells using fluorescent proteins. Exercise 8: Documentation of meristematic activity by rapid pressure method - preparation of samples. 9. Types of microscope preparations, methods of their preparation: native, permanent, whole objects and sections. Chemical and physical fixation. Exercise 9: Documentation of meristematic activity by rapid pressure method - evaluation. 10. Microscopy dyes, histochemistry and cytochemistry 11. Basics of transmission electron microscopy. Possibilities of studying cells, principles of preparation of preparations, possibilities of applications in comparison with light and fluorescence microscopy. 12. Basics of scanning electron microscopy. Possibilities of studying cells, principles of preparation of preparations, possibilities of applications in comparison with light and fluorescence microscopy.
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Learning activities and teaching methods
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Lecture, Dialogic Lecture (Discussion, Dialog, Brainstorming), Demonstration
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Learning outcomes
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Comprehensive training of students on theoretical and practical solutions of the fundamental problems of plant studies at the microscopic level
Students will acquire basic theoretical knowledge about the range of microscopic techniques and practical experiences with their applications in the study of plant cells, tissues and organs.
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Prerequisites
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unspecified
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Assessment methods and criteria
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Written exam
Practical part: 100% participation, successful completion of the credit test with a minimum of 70% of points obtained. Processing of microscopic images into a presentation (PowerPoint) according to the instructions of the trainers. Exam: Successful completion of a written exam in the form of a test with a minimum of 70% of points obtained.s
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Recommended literature
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Bargmann B.O. et al.. A map of cell type-specific auxin responses. 2013.
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Cox G. (2012). Optical imaging techniques in cell biology. CRC Press.
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Goldman, R. D., Swedlow, J. R., & Spector, D. L. (2010). Live cell imaging : a laboratory manual . Cold Spring Harbor, N.Y.
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Hejtmánek M. (1994). Praktická cvičení z biologie. Olomouc.
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Hejtmánek, M. (2001). Úvod do světelné mikroskopie. Olomouc.
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Price RL, Jerome WG. (2010). Basic confocal microscopy. Heidelberg.
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Ruzin SE . (1999). Plant microtechnique and microscopy. Oxford.
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Spector, D. L., & Goldman, R. D. (2006). Basic methods in microscopy : protocols and concepts from cells : a laboratory manual. New York, N.Y.
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Vinter V. (2011). e-learning anatomie rostlin.
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Vinter V. Rostliny pod mikroskopem (Základy anatomie cévnatých rostlin). 2009.
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Vinter V. (2006). Základy anatomie cévnatých rostlin a anatomický atlas cévnatých rostlin. Olomouc.
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