PRELIMINARY PROGRAM OF THE COURSES
General introduction to fluorescence
Introduction to fluorescent proteins (FP)
Theoretical concepts on fluorescent protein properties: folding, structure and dynamics, chromophore maturation, protonation states, oligomerization tendency, phototransformations mechanisms, photostability.
Biological function, ecological niche, diversity and evolutionary tree of FPs
Biophysical characterization of FPs in practice. FP expression and purification, optical steady state and ultrafast spectroscopy, X-ray crystallography, NMR, molecular dynamics simulations. Genetic encoding, codon optimization, fusion proteins, functional assays
Novel Fluorescent proteins
FP engineering, directed evolution, rational engineering
FP photophysics at the single molecule level, photon budget, blinking, nano-environmental effects, nonlinear light-induced FP-based sensors
Introduction to chemical dyes
Photophysics and photo-transformations, anti-fade and switching buffers, environmental influences
Advanced fluorophore photophysics: metallic environment, plasmonic resonance, nanoantennas
How to label my target in practice: labeling of proteins, DNA, lipids, cellular compartments, membranes. Halo/Snap/Clip tags, nanobodies, click chemistry. Fluorogenic probes, spontaneously blinking probes
Labeling with Q-dots and nanoparticles, lipids and sugar labeling
Labeling with expanded genetic code
Resonance energy transfer with various fluorophores and luminophores
FPs and chemical dyes: pro’s and cont’s
