- Docente: Damiano Genovese
- Crediti formativi: 6
- SSD: CHIM/03
- Lingua di insegnamento: Inglese
- Moduli: Damiano Genovese (Modulo 1) Devis Montroni (Modulo 2)
- Modalità didattica: Convenzionale - Lezioni in presenza (Modulo 1) Convenzionale - Lezioni in presenza (Modulo 2)
- Campus: Bologna
- Corso: Laurea Magistrale in Photochemistry and Molecular Materials (cod. 9074)
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Orario delle lezioni (Modulo 1)
dal 18/09/2023 al 08/01/2024
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Orario delle lezioni (Modulo 2)
dal 06/11/2023 al 09/01/2024
Conoscenze e abilità da conseguire
At the end of the course the student has acquired knowledge of the most common electronic and optical microscopies used for the characterisation of materials and biomolecules and the analysis of the acquired images.
Contenuti
The course will start with an introduction on the general concepts of digital imaging, including image formation, resolution, size, digitalization.
We will then proceed introducing the different microscopy techniques using EM waves in the range of the UV, VIS and IR:
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Optical microscopy (transmission - bright field and dark field)
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Fluorescence microscopy: the use of filters in excitation, emission and of dichroic filters. Detection cameras
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Confocal microscopy: the pinhole, the scanner unit, signal multiplexing and decoding. Point and line measurements, dynamic measurements, FCS and correlative microscopies. Multiplexing in wavelength (channels vs spectral module) and in time (FLIM and PLIM, fluorescence and phosphorescence lifetime imaging). Dynamic techniques (FRAP, FLIP).
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Multiphoton microscopy
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Special configurations: light sheet microscopy
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Beyond diffraction limit: super-resolution microscopies
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Brief in-depth information on fluorescent probes that meet specific microscopy needs (brightness, super-resolution, photostability…)
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Brief introduction to Raman and IR microscopies
The second course section introduces the students to the principles and applications of imaging technologies based on electron microscopy (EM). Different microscopy techniques will be described: scanning (SEM), transmission (TEM), scanning transmission (STEM), etc.
The class will also discuss techniques that can be combined to EM to get additional information on the sample, i.e., composition or crystallinity. These include energy-dispersive X-ray spectroscopy (EDS), cathodoluminescence (CL), electron backscatter diffraction (EBSD), focused ion beam (FIB), etc.
A brief and not in-depth introduction to scanning microscopies (AFM, SECM, etc.) will also be given to introduce this class of techniques and their range of application compared to EMs.
The course will discuss the technique and theoretical bases behind each of these classes of EMs along with their limitations and range of application. Aside of the basic theory, a strong relevance will be given to practical knowledge as sample preparation and high image quality acquisition.
Finally, the student will learn the basics of image analysis to perform quantitative experiments and measurements.
Testi/Bibliografia
Single-molecule optical detection, imaging and spectroscopy / edited by T. Basche
Weinheim: VCH, c1997
Fluorescence spectroscopy, imaging and probes : new tools in chemical, physical and life sciences / Ruud Kraayenhof, Antonie J. W. G. Visser, Hans C. Gerritsen (eds.)
Springer, c2002
Bellingham : Spie press, c2006
Metodi didattici
Lezioni Frontali con sessioni pratiche
Modalità di verifica e valutazione dell'apprendimento
Esame Orale - Fac-simile di una sessione sperimentale di microscopia
Strumenti a supporto della didattica
Slides Powerpoint saranno rese disponibili su Virtuale
Orario di ricevimento
Consulta il sito web di Damiano Genovese
Consulta il sito web di Devis Montroni