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Lecturer(s)
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Doležel Jaroslav, prof. Ing. DrSc.
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Course content
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Classification of cytometric techniques, cytomics. Flow cytometry, history, parameters measurable using flow cytometry, basic components of a flow cytometer. Fluidics, laminar flow, flow chamber, hydrodynamic focusing. Flow sorters and their construction designs, application of flow cytometric sorting. Light sources, arc lamps, lasers, optical filters. Detection of light pulses, photodiodes and photomultipliers, parameters of a light pulse. Signal amplifiers, digital signal processing, fluorescence compensation, presentation of results of analysis. Application of flow cytometry in plants, analysis and sorting of mitotic chromosomes, protoplast analysis, analysis of chloroplasts, analysis of gene expression, study of phytoplankton, applications in phytopathology, Luminex technology. Estimation of nuclear DMA content, determination of ploidy, estimation of genom size, identification of interspecific hybrids, detection of aneuploids, determination of mode of reproduction, analysis of cell cycle kinetics, apoptosis, endoreduplication, sorting of cell nuclei. Flow cytometry in biomedicine. Immunophenotyping, CD nomenclature, mitochondria and apoptosis, mitochondrial membrane potential, fluorescence resonance energy transfer (FRET).
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Learning activities and teaching methods
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Lecture, Monologic Lecture(Interpretation, Training), Dialogic Lecture (Discussion, Dialog, Brainstorming)
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Learning outcomes
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View into cytometric technuques, their principles and aplication in plant genetics and biomedicine.
Student is able to (after attending the course): - Explain the principles of flow cytometry. - Describe the function of individual components of the instrument. - Apply flow cytometry in plant sciences research. - Discuss the use of flow cytometry in biomedical research and clinical practice. - Judge the usefulness of the method to solve a particular problem. - Interpret the results obtained using flow cytometry. - Evaluate critically research papers.
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Prerequisites
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unspecified
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Assessment methods and criteria
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Oral exam
75% attendance, oral colloquium
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Recommended literature
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Darzynkiewicz, Z., Crissman, H.A. (1990). Methods In Cell Biology: Flow Cytometry. Academic Press, San Diego.
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DARZYNKIEWICZ, Z., CRISSMAN, H.A., ROBINSON, J.P. (2000). Methods in Cell Biology: Cytometry, 3rd Edition. Academic Press, San Diego.
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DOLEŽEL, J., GREILHUBER, J., SUDA, J. (2007). Cytometry with Plant Cells. Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
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Eckschlager, T. a kolektiv. (1999). Průtoková cytometrie v klinické praxi. Grada Publishing, Praha.
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Robinson, J.P., Darzynkiewicz, Z. (1997). Current Protocols in Cytometry. John Wiley & Sons, New York.
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Shapiro, H.M. (2003). Practical Flow Cytometry, 4th Edition. John Wiley and Sons, Inc., New York.
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