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Optické spektroskopie 1

Course: Optical Spectroscopies 1

Department/Abbreviation: KEF/OSP1

Year: 2021 2022

Guarantee: 'doc. RNDr. Martin Kubala, Ph.D.'


  • Sources of light and ways of excitation. Absorption spectroscopy. Experimental technique of absorption spectroscopy. Luminescence. Applications of fluorescent spectroscopy. Fluorophores.

    Course review:
    1. Introduction (spectroscopies - what is it and why do we use it, interaction of light and matter, energy of light, units, UV/VIS/NIR spectral range). 2. Sources and detectors of light (lasers, diodes, lamps, synchrotron rays, CW-, pulsed- and modulated-light, total internal reflection, principles of light detection, photomultipliers, photodiodes, CCD). 3. Absorption spectroscopy (derivation of Lambert-Beer law, presentation of spectra, photobleaching, transient absorption, experimental setup in absorption spectroscopy, experimental technic in absorption spectrocopy). 4. Absorption of biologically important molecules, perception of light, detection of light by living organisms. 5. Luminescence. Phenomenon of luminescence, Jabloński diagram, sorting of luminescences, sorting of photoluminescences, delayed fluorescence, elementary characteristics of luminescence. 6. Experimental technic for measurement of photoluminescence. Experimental setup in luminescence spectroscopy (comparison to absorption s.), spectral limits, sources, monochromators, samples, detectors, other optical elements. 7. Steady-state fluorescence (intensity, excitation-, emission and synchronnous spectra, quantum yield). 8.Fluorescence measurement using a pulsed light (time-domain) (kinetic, DAS, TRES). 9. Fluorescence measurement using a harmonically-modulated light (phase-domain). 10. Fluorophores. Intrinsic fluorophores, fluorescence probes, sensors, fluorescent proteins.10. Single-molecule spectroscopy. 11. Measurements using polarized light (anisotropy, rotational correlation time, Perrin equation, time-resolved measurement). 12. Fluorescence quenching (static, dynamic, Stern-Volmer equation). 13. FRET. 14. Fluorescence microscopy (confocal, FLIM, STED). 15. Measurement of intensity fluctuations (FIDA, FCS, FRAP). 16. Single-molecule spectroscopies. 17. Multiphoton excitation of fluorescence. 18. Solvent effects.


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    Videozáznamy přednášek:

    13 FRET (MP4 106 MiB)

    14 Fluorescenční mikroskopie (MP4 71 MiB)

    15 Měření fluktuací intenzity (MP4 39 MiB)

    16 Spektroskopie jednotlivých molekul (MP4 31 MiB)

    17 Multifotonová excitace fluorescence (MP4 21 MiB)

    18 Vliv solventu (MP4 38 MiB)


    Otázky ke zkoušce (PDF)345.61 KB
    Výsledky testů (DOC)79 KB