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Lecturer(s)
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Stiborová Kateřina, Mgr.
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Fluks Monika, Ph.D.
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Hauserová Andrea, Mgr.
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Matyášová Alena, Bc.
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Žurovec Michal, prof. RNDr. CSc.
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Sehadová Hana, Mgr. Ph.D.
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Weyda František, doc. RNDr. CSc.
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Bruce Alexander William, prof. Ph.D.
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Podlahová Šárka, Mgr.
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Rouhová Lenka, Mgr.
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Brož Václav, Mgr. Ph.D.
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Course content
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Presentations: 1. Histology (treatment, fixation, and staining of tissues, paraplast sections, total preparations) 2. Detection of proteins on the tissue and cellular level (immunohistochemistry, practical achievements, biopsy) 3. Detection of DNA, RNA on the tissue and cellular level (in situ hybridization, practical achievements, FISH) 4. Bright-field microscopy (background + technique for contrast increasing) 5. Fluorescence and confocal microscopy 6. Electron microscopy 7. Isolation of DNA, RNA and proteins 8. PCR, real-time PCR, microarrays and their practical achievements in diagnostics 9. Sequencing and basics of transcriptomics 10. Tissue cultures 11. Embryogenesis of mammals (gametogenesis, fertilization and cleavage of zygote, artificial fertilization, development of tweens) Practical trainings: 1. Preparation of the paraplast and semithin sections 2. Histological staining - Mallory, hematoxylin-eosin 3. Immunohistochemical detection of specific proteins in the insect tissues 4. Giemsa staining of blood smears, determination of blood group based on clinical tests 5. Training on bright-field, fluoresce and confocal microscopes 6. Excursion to the laboratory of the electron microscopy 7. DNA isolation, gel electrophoresis 8. PCR, real-time PCR, and gel electrophoresis 9. Excursion to the clinical pathology (visit of histological laboratory, biopsy) Observation of cleavage of the mouse zygote 10. Tests of cells viability - efficiency of drugs 11. Isolation of oocytes from mouse ovaria, activation of meiosis
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing), Dialogic (discussion, interview, brainstorming), Monitoring, Demonstration, Laboratory, Excursion, Work with multi-media resources (texts, internet, IT technologies), Experiment, Practical training
- Class attendance
- 84 hours per semester
- Preparation for classes
- 40 hours per semester
- Preparation for credit
- 40 hours per semester
- Field trip
- 20 hours per semester
- Preparation for exam
- 40 hours per semester
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Learning outcomes
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Aim of course is to present recent classic as well as completely new methods of cell research. Emphasis is laid to dynamic aspects of basic techniques used in optical, confocal and conventional electron microscoy. Special attention is directed to correlative microscopy and determination of artefacts.
Students meet on theoretical as well as on practical level with broad spectrum of methods used especially in cell biology
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Prerequisites
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The course introduces the issue, does not assume specific input knowledge.
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Assessment methods and criteria
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Oral examination, Student performance assessment, Analysis of student's work activities (technical works), Test, Development of laboratory protocols
Requirements for credit: 1) presence on the practical trainings (maximum one absence) 2) submission of protocols from each practical training 3) successful passing of final test Requirements for oral exam: 1) obtaining of the credit
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Recommended literature
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Celis J.E. (ed), 1994: Cell Biology. A Laboratory Handbook. Academic Press (San Diego), Vol.1-3.
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Hunter E., 1993: Practical Electron Microscopy. Cambridge University Press (Cambridge), 173 stran.
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Pollak J.M., Priestley J.V (eds), 1992: Electron Microscopic Imunocytochemistry. Oxford University Press (Oxford), 267 stran.
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Williams D.B., Carter C.B. 1996: Transmission Electron Microscopy. Plenum Press (New York), Vol.I-IV.
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Heath Julian P. Dictionary of Microscopy. 2000. ISBN 0-470-01199-8.
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Kuo, John (Ed.). Electron Microscopy. Methods and Protocols.. 2014. ISBN 978-1-62703-776-1.
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S. Amelinckx (Editor), Dirk van Dyck (Editor), J. van Landuyt (Editor), Gustaaf van Tendeloo (Editor). Electron Microscopy: Principles and Fundamentals.. 2008. ISBN 978-3-527-61455-4.
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