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Lecturer(s)
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Vancová Marie, RNDr. Ph.D.
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Kaftan David, Mgr. Ph.D.
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Gardian Zdenko, Ing. Ph.D.
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Bílý Tomáš, Mgr. Ph.D.
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Course content
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Lectures: The visualisation of biological macromolecules in high resolution. Qualitative and quantitative analysis of biological objects in electron microscopes. Progressive methods of biological specimen preparation. Advanced methods of immunolabeling. 3D electron microscopy. Electron tomography: from sample preparation to 3D visualization of CLEM = correlative light and electron microscopy. Probe microscopies (AFM, STM) and their biology applications. Practical: 3D reconstruction of the biological specimen using the method of electron tomography Single particle analysis of negatively stained samples Cryo-cutting and immunolabeling Demonstration of AFM
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing), Dialogic (discussion, interview, brainstorming), Laboratory
- Preparation for credit
- 24 hours per semester
- Preparation for classes
- 28 hours per semester
- Preparation for exam
- 24 hours per semester
- Class attendance
- 24 hours per semester
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Learning outcomes
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Lectures: Immunoelectron microscopy with unlabelled and labeled antibodies and their applications. Cryometries in Biological Electron Microscopy. Electronic tomography. Visualization of biological macromolecules. Autoradiography. Image analysis, three-dimensional reconstruction in electron microscopy. Morphometry and stereology. Electronic microanalysis (quality, quantity) of biological objects. Correlative microscopy. Electronic diffraction and its application in biology. New microscopy (STM, AFM) and their biological applications. A Lab exercise: Preparation of ultra-thin sections, sectional immunoassay, identification of crystalline structure by electron diffraction, determination of particle size in suspension by means of SEM, processing of measurement in ACC program. Překladač Google pro firmy:Translator ToolkitPřekladač webových stránek
Students receive an overview of advanced methods of electron and probe microscopy and knowledge of their use in biology and biolmedical research. Students acquire basic knowledge about the preparation of samples for those advanced methods. Students learn about cellular ultra-structure, and get an idea of the the possibilty to detect biomolecules. Students learn about the possibility of using electron microscopes for quantitative and qualitative electron microanalysis.
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Prerequisites
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Exam from the course "Electron Microscopy I"
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Assessment methods and criteria
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Test
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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Jana Nebesářová. Elektronová mikroskopie pro biology. Č.Budějovice, 2002.
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John Kuo. Electron Microscopy. Springer Heidelberg, 2014. ISBN 978-1-62703-776-1.
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