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Lecturer(s)
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Ďurinová Eva, Mgr.
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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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Kitzberger František, Mgr. MSc.
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Course content
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Content of lectures: ¨ Physical Principles of Electron Microscopy, Physical Properties of Accelerated Electrons, Resolution, Magnification. Transmission electron microscope, electromagnetic lenses and their defects, device design. Specimen preparation for TEM by physical methods, cryo-mmethods, using microwaves. Specimen preparation for TEM by chemical methods, fixation, dehydration, embedding. Preparation of ultrathin sections, ultramicrotome, knives, specimen contrasting, supporting foils. Immunolocalisation methods. Scanning electron microscopy, device design. Specimen preparation for SEM, drying and coating methods. Electron-microscopic cytology. 9. Artifacts in TEM and SEM caused by specimen preparation, device adjustment, recording process 10. Recording of digital image in TEM and SEM. Methods of 3D electron microscopy. Practical exercises: Preparation of a biological object for TEM - methods of fixation, dehydration and embedding. Trimming procedure, cutting of semi-thin and ultrathin section using glass knives, staining of sections. Preparation of a plant virus suspension using the method of negative staining. Specimen preparation for SEM at RT. Method of critical point drying, method of sputter coating. Adjustment and basic control of TEM and SEM, examination of prepared specimen in SEM and TEM. Digital image recording in SEM and TEM and the basics of image processing. Content of practices:
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing), Dialogic (discussion, interview, brainstorming), Laboratory, E-learning
- Class attendance
- 60 hours per semester
- Preparation for credit
- 24 hours per semester
- Preparation for exam
- 60 hours per semester
- Preparation for classes
- 56 hours per semester
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Learning outcomes
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To acquaint students with basic physical principles of transmission and scanning electron microscopy and methods of preparation of biological objects for electron microscopy.
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 that students have 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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Combined exam
Participation in the laboratory part of the course is obligatory. Prior to every laboratory exercise, the student shall take a test proving their theoretical knowledge.
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Recommended literature
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