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Laboratorio di Biofisica

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Anno accademico 2008/2009

Codice dell'attività didattica
8049S
Docente
Prof. Renzo Levi (Titolare del corso)
Corso di studi
c210 laurea spec. in fisica ambientale e biomedica
Anno
2° anno
Tipologia
Di sede o curricolari
Crediti/Valenza
3
SSD dell'attività didattica
BIO/09 - fisiologia
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Sommario insegnamento

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Programma

Introduzione all'uso delle tecniche di fluorescenza, alla microscopia confocale
Introduzione alle tecniche di image processing e image analysis

Physical techniques in biological research have a continuous development.
Among these techniques the laboratory course focuses on optical techniques based on fluorescent probes, on image processing of fluorescence images and on advanced electrophysiological
cell techniques based on the patch-clamp approach.

What is fluorescence? The Jablonski diagram, molecular bases, timescales of the events, environment dependence of fluorescence.
Related processes, quenching, FRET...
Organic fluorescent probes, quantum dots and engineered fluorescent proteins: their application domains.
Tools to evaluate fluorescence and their main components. Light sources and their spectra, dichroic filters, microscope lenses,
and light detectors (PMT, CCD and others).
Principles of confocal microscopes, with an example of image acquisition in 3d microscopy.
Image processing and image analysis, a hands on approach using ImageJ.

A practical approach to single cell electrophysiology analyzing the components of a patch-clamp setup and measuring
membrane potential and ionic currents on a neural derived cell line.
Components of the system: the glass electrodes and the conducting solutions, the electronics, the mechanical
components (manipulators and antivibration table). The data acquisition and analysis platform.
Examples of ionic currents evoked by different voltage protocols.

Testi consigliati e bibliografia



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