Lecture
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18 Nov 2025 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Experimental uncertainties, probability distributions (Gaussian, Lorentzian, Poissonian), error bars, hints on statistical tests; statistical estimators (average, standard deviation, standard error, confidence level). Error propagation. Covariance.
21 Nov 2025 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Covariance matrix and correlations. [Nota: interruzione per allarme antincendio] Noise in measurements, examples in spectroscopy. Microscopy: fundamentals of lenses; objectives, resolution, numerical aperture. Electromagnetic radiation, refraction and diffraction of light, limit of diffraction for resolution (Rayleigh criterium).
25 Nov 2025 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Additional considerations on resolution; Gaussian beams. Optical path in transmission and reflection/epifluorescence microscopes, conjugate planes, field and aperture diaphragms. Köhler illumination. Contrast definitions and techniques (introduction, polarization contrast, dark field, phase contrast, DIC); optical path and phase differences. Alignment of a transmission microscope. Contrast techniques in reflection microscopy. Epifluorescence microscopy. Fluorophores and immunofluorescence.
02 Dic 2025 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Dichroics and optical filters. Confocal microscopy: Introduction, implementations, Point Spread Function, convolution; magnification and resolution; dependence on signal/noise ratio, variable with laser intensity and detector gain.
05 Dic 2025 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Noise in microscopy, effect of accumulations/averages. Dependence of signal/noise ratio and of resolution on the confocal pinhole aperture. Aberrations (spherical, chromatic). 2 photon microscopy (introduction). Two photon microscopy: implementation (pulsed lasers); TIRF microscopy: dipole emission, distortions in TIRF.
09 Dic 2025 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Fluorescence microscopy: labelling (fluorescent proteins, Qdots, organic fluorophores). Metals, semiconductors, insulators. Chemical tags: methods of insertion into proteins (transfection, vectors, plasmids, promoters). Lentiviral vectors, promoters, controlled expression, insertional mutagenesis, hints on PCR, primers. Advanced methods: colocalization and its quantifications. FRET: applications; efficiency measurement by acceptor photobleaching, calculations of sensitized emission corrections.
12 Dic 2025 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
FLIM (hints). FRET-FLIM with examples. Laws of diffusion, sum over microtrajectories, Stokes-Einstein equation; solution of the diffusion equation starting with a Dirac Delta for the concentration. FRAP: experimental details (corrections, normalizations, geometries, bleaching, examples). Analysis of FRAP: diffusion model.
19 Dic 2025 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Analysis of FRAP: diffusive model, compartment model. Other "FRAP-like" microscopy techniques based on photobleaching: FLIP; iFRAP, photoactivation (PA), FLAP, photoconversion (PC). FCS: (auto)correlation function, its fit for diffusion phenomena, other contributions (dark states, molecules rotation, antibunching). Spectroscopy of single molecules (outline and techniques).
09 Gen 2026 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Single molecules tracking: analysis; types of trajectories, MSD; anomalous diffusion, transient arrest of diffusion. Hints on experiments on membrane receptors and neurotrophins: 2D-distributions of couples of parameters, counting fluorescent molecules in tracked spots. Anomalous diffusion by transient confinement zones and by diffusion-reaction; log-log graphs and power dependencies. Deconvolution. Structure of biological molecules: molecular forces and energies.
13 Gen 2026 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Covalenti and non-covalent forces. Hydrophobic effect. Carbohydrates: stereochemistry, isomerism; structures and conformations of monosaccharides; glycosidic bonds, disaccharides, polysaccharides (cellulose, starch, glycogen). Glycoproteins and glycolipids. Lipids: categories; fatty acids, triglycerides; membrane lipids. Plasma membrane structure. Nucleic acids (NA): DNA and RNA (structures, functions). Artificial NA. Polymerization and hydrolysis; interactions amongst bases; different helices.
16 Gen 2026 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Nucleic acids (NA): different helices, persistence length; nucleosomes, chromatin, additional secondary/tertiary structures. Proteins: amino acids, peptide bond; posttranslational modifications; dihedral angles and Ramachandran plots; secondary structures: helices, beta sheets, turns, loops; tertiary structure and classification; quaternary structure; protein folding, stability and denaturation, flexibility and function. Experimental determination of structures. Protein crystals production.
23 Gen 2026 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Folding of proteins: simulations. Experimental determination of structures: x-ray cristallography, missing phase problem and solutions, resolution. Protein data bank and PDB files. Structure determination by NMR; hints on different methods. Light-matter interactions: introduction, generalities. Quantum mechanics recall: quantum states, vectorial spaces.
27 Gen 2026 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Quantum Mechanics: operators, eigenstates and eigenvalues; Schrödinger equations; solutions for constant potential, for step-like potentials, quantum wells. Time-dependent perturbation theory, Fermi's golden rule. Review of Fourier series and transform, Taylor series; Dirac delta.
03 Feb 2026 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Fermi Golden Rule: recall. Interactions with radiation. Time evolution of combination of two stationary states. Einstein coefficients and black-body radiation. Operating principles of lasers. Jablonski diagrams and absorption. Electronic and vibrational spectroscopy: Born Oppenheimer approximation, IR absorption. Vibrational modes and selection rules.
06 Feb 2026 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Vibronic transitions; Franck Condon factors. Properties, setups and measurement methods for optical absorption (UV-visible-NIR); spectrometers and grating. Lambert-Beer law. Properties of fluorescence and phosphorence. Fluorescence: the three properties; measurements methods and setups. Spectrofluorometer interface.
10 Feb 2026 (2h 00m)
STEFANO Luin - Corso (attività didattica) - Mista
Brightness. Non-linear light-matter interactions. Differences amongst 2-photon absorption and second harmonic generation. Second order perturbation theory, 2 photon absorption, Raman. Raman: Stokes/antiStokes intensity ratio, thermally activated fluorescence. Vibrational spectroscopy: typical energies, vibrations localized on bonds. Organic fluorophores: orbitals; molecular transitions. Molecular states of formaldehyde (introduction).
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