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Lecturer(s)
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Vancová Marie, RNDr. Ph.D.
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Bílý Tomáš, Mgr. Ph.D.
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Gardian Zdenko, Ing. Ph.D.
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Týč Jiří, RNDr. Ph.D.
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Ďurinová Eva, Mgr.
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Kitzberger František, Mgr. MSc.
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Course content
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Lectures: 1. Physical Principles of Electron Microscopy, Physical Properties of Accelerated Electrons, Resolution, Magnification 2. Transmission electron microscope, electromagnetic lenses and their defects, device design 3. Specimen preparation for TEM by physical methods, cryo-mmethods, using microwaves 4. Specimen preparation for TEM by chemical methods, fixation, dehydration, embedding. 5. Preparation of ultrathin sections, ultramicrotome, knives, specimen contrasting, supporting foils 6. Scanning electron microscopy, device design 7. Specimen preparation for SEM, drying and coating methods 8. Electron-microscopic cytology 9. Artifacts in TEM and SEM caused by specimen preparation, device adjustment, recording process 10. Digital and photographic recording of a microscopic image Practical exercises: Preparation of semi-thin sections, treatment of microscopic grids and oating of grids by formwar layer, visualization of the plant virus using the method of negative contrast, specimen preparation for SEM, operation with TEM and SEM and examination of samples prepared by the method of negative staining
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing), Dialogic (discussion, interview, brainstorming), Laboratory
- Class attendance
- 48 hours per semester
- Preparation for credit
- 16 hours per semester
- Preparation for classes
- 16 hours per semester
- Preparation for exam
- 20 hours per semester
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Learning outcomes
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1. Physical principles of electron microscopy, physical properties of accelerated electrons, resolution, magnification 2. Transmission electron microscope, electromagnetic lenses and their defects, device design 3. Preparation of preparations for TEM by physical methods, cryometries, microwaves 4. Preparation of preparations for TEM by chemical methods, fixation, dehydration, watering 5. Preparation of ultraviolet cuts, ultramicros, knives, contrast, backing foil 6. Scanning electron microscopy, device design 7. Preparation of SEM preparations, drying, plating 8. Electromicroscopic cytology 9. Artifacts in TEM and SEM due to preparation, apparatus, photographic process 10. Digital and photographic recording of a microscopic image A Lab exercise: Preparation of half-cuts, mesh trimming and molding membrane coating, visualization of plant virus method of negative contrast, preparation of SEM preparation itself, work with TEM and SEM and inspection of prepared preparations
Students receive an overview of highest resolution microscopic methods and knowledge of their use in biology and biolmedical research. Students acquire basic knowledge about the preparation of samples by chemical and physical techniques. Students learn about cellular ultra-structure, and get an idea of the structure, appearance and size of basic cellular organelles.
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Prerequisites
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The course assumes the knowledge of physics, chemistry and biochemistry at the level of completed secondary education
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Assessment methods and criteria
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Written examination
Elaboration of protocols from selected laboratory exercises. 60% of correct answers in the final written exam.
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Recommended literature
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MJ Dykstra, LE Reuss: Biological Electron Microscopy ? Theory, Techniques and Troubleshooting. 2nd Edition..
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