Syllabus
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
Annotation
Methods of optical absorption spectroscopy, light scattering and Raman spectroscopy, chiroptical and luminescence methods as applied in biophysical research.