The course will provide advanced knowledge of microscopic techniques and their possible applications in the biological field, and in particular in basic and applied biomedical research.

COURSE TOPICS
Light microscopy: operating principles and applications. The cyto-histological techniques as a diagnostic method. Histochemical methods. Super resolution microscopy. Laser microdissection microscope. Microinjection. Immunohistochemical and immunofluorescence procedures: application in biological field and applied biomedical research. Antibodies and immunostaining. Fluorescence and fluorochromes. Fluorescent proteins. The fluorescence microscopy and confocal laser scanning microscopy: guiding principles of specimen preservation for confocal fluorescence microscopy. Two-photon Laser Scanning Microscopy. In situ hydridization: principles and applications. The transmission and scanning electron microscopy: specimens preparation and observation. The Value of Electron Microscopy in Diagnosis. Immunogold labelling and applications in biomedical field. Microanalysis element identification: preparation and observation of samples. Evaluation of the presence of substances in biological samples. The environmental scanning electron microscope and applications. Immunogold labelling in environmental scanning electron microscopy: applicative features for complementary cytological interpretation. Microscopy in nanotechnology. Atomic Force Microscopy: applications in pharmaceutical research. Image analysis. High Content Analysis. Discussion of scientific papers.

Description of the practical activities.
Preparation of histological slides: fixation, inclusion, paraffin section preparation, staining of samples and observation with optical microscope. Immunohistochemical method: the reaction ABC immunoperoxidase. Sample preparation for fluorescence microscopy: observation under a fluorescence microscope and confocal laser scanning microscope. Sample preparation for transmission electron microscopy: fixation, dehydration, resin embedding, thin sectioning, negative and positive staining. TEM observation. The immunogold labelling: sample preparation and observation. The scanning electron microscope: sample preparation and observation. Elemental microanalysis: sample preparation and observation.
The practical activities will be carried out at the Zoology and CGA laboratories.

Slides and scientific articles provided by the teacher.