Fundamentals of Biophysics at the Nanoscale

Lecture log

Academic year 2025/2026
Lecturer Francesco Cardarelli, Stefano Luin, Gian Michele Ratto

Lecture

  • 18 Nov 2025 (2h 00m)

    STEFANO Luin - Course (teaching activity) - Both face to face and online

    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 - Course (teaching activity) - Both face to face and online

    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 - Course (teaching activity) - Both face to face and online

    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 Dec 2025 (2h 00m)

    STEFANO Luin - Course (teaching activity) - Both face to face and online

    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 Dec 2025 (2h 00m)

    STEFANO Luin - Course (teaching activity) - Both face to face and online

    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 Dec 2025 (2h 00m)

    STEFANO Luin - Course (teaching activity) - Both face to face and online

    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 Dec 2025 (2h 00m)

    STEFANO Luin - Course (teaching activity) - Both face to face and online

    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.