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Optical Spectroscopy Methods in Biophysics - NBCM113
Title: Metody optické spektroskopie v biofyzice
Guaranteed by: Institute of Physics of Charles University (32-FUUK)
Faculty: Faculty of Mathematics and Physics
Actual: from 2020
Semester: winter
E-Credits: 5
Hours per week, examination: winter s.:4/0, Ex [HT]
Capacity: unlimited
Min. number of students: unlimited
4EU+: no
Virtual mobility / capacity: no
State of the course: taught
Language: Czech
Teaching methods: full-time
Teaching methods: full-time
Guarantor: prof. RNDr. Josef Štěpánek, CSc.
prof. RNDr. Petr Heřman, CSc.
Classification: Physics > Biophysics and Chemical Physics
Interchangeability : NBCM177
Is incompatible with: NOOE004
Annotation -
Last update: G_F (07.01.2003)
Methods of optical absorption spectroscopy, light scattering and Raman spectroscopy, chiroptical and luminescence methods as applied in biophysical research.
Aim of the course -
Last update: STEPJOS/MFF.CUNI.CZ (15.05.2008)

To learn students theoretical and technical basics of molecular optical

spectroscopy and its application in study of biological targets.

Course completion requirements - Czech
Last update: prof. RNDr. Josef Štěpánek, CSc. (30.10.2019)

Oral exam.

Literature -
Last update: STEPJOS/MFF.CUNI.CZ (15.05.2008)

O.Jelínek,P.Pančoška,J.Štěpánek: Metody optické spektroskopie (v uč. \"Experimentální metody

biofyziky\",V.Prosser a kol.), Academia, Praha 1989

Parker C.A.: Photoluminescence of Solutions, Elsevier, Amsterdam (1986) - ruský překlad

Fotoljuminescencija rastvorov, Izd. nauka, Moskva (1972)

Vacek K., Pantoflíček J., Jelínek O.: Luminiscence I a II, SPN Praha (1972)

Jelínek O.: Experimentální metody biofyziky III - Luminiscence, SPN Praha (1983)

Lakowicz J.R.: Principles of Fluorescence Spectroscopy, Plenum Press, New York, second edition (1999)

Alfano E.R. (ed.): Biological Events Probed by Ultrafast Laser, Spectroscopy, Acad. Press, New York-

London (1982)

Žaloudek F., Plášek J., Sladký P.: Vybrané metody optické spektroskopie, SPN Praha (1982)

Nepraš M., Titz M.: Základy teorie elektronových spekter , SNTL Praha (1983).

Teaching methods -
Last update: STEPJOS/MFF.CUNI.CZ (15.05.2008)

lecture

Requirements to the exam - Czech
Last update: prof. RNDr. Josef Štěpánek, CSc. (16.10.2017)

Zkouška je ústní. Požadavky ke zkoušce jsou v rozsahu zveřejněného sylabu.

Syllabus -
Last update: T_FUUK (11.05.2004)

Conversion of the excitation energy to luminescence - general rules: Jabloňski diagram, radiative and nonradiative transitions, fluorescence and phosphorescence, internal conversion and intersystem crossing

Basic characteristics of fluorescence: intensity, excitation and emission spectra, quantum yield, fluorescence lifetime and its relation to absorption, polarized fluorescence in solution, fluorescence anisotropy, polarization and anisotropy terms, photoselection, Perrin equation

Measurements of the steady-state spectra: basic fluorimeter set-up, excitation light sources, filters and polarizers, monochromator, analog detection and photon counting, correction of emission spectra, quantum yield measurements

Fluorescence lifetime measurements: time domain: fluorescence intensity decay, response function, convolution integral, magic angle, average lifetime, time-correlated single-photon counting method, streak camera; frequency domain: basic principles, modulation and phase dependence for single lifetime decay, frequency domain fluorimeter set-up

Solvent effects on emission spectra: general effect, Lippert equation, specific interactions, temperature effects

Fluorescence quenching: dynamic quenching, collisional quenching, Stern-Volmer equation, static quenching, applications of quenching to proteins

Fluorescence energy transfer: transfer rate from donor to acceptor, Főrster distance, overlap integral, orientation factor, molecular distance measurements

Excimer fluorescence: excimer, exciplex, two-state model, basic spectral and lifetime characteristics

Fluorophores: intrinsic or natural, extrinsic, protein fluorescence, fluorescent probes

 
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