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Lecturer(s)
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Kopecký Vladimír, RNDr. Ph.D.
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Bína David, RNDr. Ph.D.
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Litvín Radek, RNDr. Ph.D.
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Course content
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Content of lectures: 1. Theoretical background: electromagnetic radiation, interaction of light with matter, absorption, emission, dispersion 2. Practical aspects: radiation sources, detection methods; principles, and use 3. Methods based on dispersion of light by macromolecules, DLS; absorption spectroscopy part I 4. Absorption spectroscopy part II: time resolved methods 5. Linear and circular dichroism 6. Fluorescence I: introduction to the method, excitation and emission spectra, fluorophores, FRET 7. Fluorescence II: time resolved fluorescence, fluorescence correlation spectroscopy 8. Optical microscopy: confocal microscopy, localization, superresolution methods 9. Rotational-vibrational spectroscopy: introduction 10. FTIR 11. Methods of Raman spectroscopy, resonance Raman spectroscopy 12. Theoretical practicals to summarise covered topics, example solution of selected problems by using optical spectroscopy methods Content of practicals: In practicals, the information from the lectures will be covered in more detail, methods will be demonstrated.
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing), Dialogic (discussion, interview, brainstorming), Excursion, Practical training
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Learning outcomes
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The course will present theoretical and practical information on optical experimental methods and their use in biochemistry
The students will acquire understanding of the theoretical background of absorption and emission of light. The course covers principles, advantages, disadvantages and important experimental aspects of covered methods of optical spectroscopy and imaging. A large part of the covered methods will be demonstrated on biochemically relevant samples.
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Prerequisites
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The students should understand introductory topics of light as electromagnetic radiation and its properties. The desired level is about that of advanced high school physics curriculum.
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Assessment methods and criteria
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Combined exam
Correct answers to at least 50% of exam questions.
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Recommended literature
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Lakowicz J.R.: Principles of fluorescence spectroscopy, 3rd edition, Springer, 2006.
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Prosser V., ed.: Experimentální metody biofyziky, Academia, 1989.
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Selvin P.R., Ha T.: Single-molecule techniques, CSH Press, New York, 2008.
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van Holde K.E., Johnson W.C., Ho P.S.: Principles of Physical Biochemistry, 2nd edition, Prentice Hall, Upper Saddle River, NJ, 2005.
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