Integrative teaching
Francesco Cardarelli, Ciclo Di Seminari
Exercises
Examination procedure
Oral exam
Prerequisites
Recommended for PhD students in Nanosciences, suggested for PhD students in Neurosciences, Methods and Models for Molecular Sciences, indicated also for PhD students in Physics.
Prerequisites: basic principles of Optics, of organic and inorganic Chemistry, and of Quantum Mechanics.
Syllabus
Measurements in microscopy and spectroscopy [1-4]
Noise in measurements, experimental uncertainties, basics of probability distributions, propagation of uncertainties.
Transmission, reflection and epifluorescence microscopy.
Magnification and resolution; contrast techniques; spherical and chromatic aberrations; hints on optical filters and dichroics.
Confocal microscopy: set-up, point spread function, hints on deconvolution, comparison with TIRF and 2-photon microscopy.
Light-matter interaction: fundamentals (also quantum mechanics) and setups for absorption, fluorescence, Raman, and multiphoton excitation. Jablonski diagrams and properties of fluorescence. Organic dyes: chemical structures and exploitation in fluorescence microscopy.
Hints on fluorescent quantum dots. Fluorescent proteins, GFP family.
Diffusion and Brownian motion. Techniques in fluorescence microscopy: colocalization, FRAP-like techniques, FRET, FLIM (fundamentals, instruments, phasors), FCS, super-resolution (RESOLFT, STED, F-PALM, SIM), single molecule spectroscopy and tracking.
Basis of molecular and cellular biology [5]
See supplementary teaching
Introduction to the structure and dynamics of biological molecules [6]
Fundamentals of electrophysiology and in-vivo microscopy
See cycles of seminars
Bibliographical references
[1] "An Introduction to Error Analysis", J. R. Taylor (Ch. 1-4, 9, 11)
[2] "Microscopy from the very beginning", Dr. H. G. Kapitza, © Carl Zeiss Jena GmbH, 1997, 2nd revised edition, on-line available
[3] "Introduction to Confocal Fluorescence Microscopy", Michiel Müller, edited by SPIE press (WA, USA), second edition (2006)
[4] "Fluorescence Applications in Biotechnology and Life Sciences", edited by Ewa M. Goldys (2009), published by John Wiley & Sons (Hoboken, NJ, USA), Ch. 1-6, 9-11, 16.
[5] "Molecular Biology of the cells", B. Alberts et al. (chosen parts)
[6] "Biophysical Chemistry", Cantor and Schlimmel; Part I