Lesson: Overview of RNAse-free treatments, probe synthesis and in situ hybridization techniques (tissue fixation, hybridization, immunolocalization of probes, double labelling).

Practice: Students will focus their practical experience on RNA probes synthesis and whole mount in situ hybridization using animal model systems. Students will fix animals and will hybridize them using different cell/tissue specific RNA probes. In particular, students will use probes specific for stem cells, nervous system, eyes and gut.

Lesson: The potential value of transmission electron microscopy (TEM) in life sciences. Short history of electron microscopy in Italy. Overview on basic techniques in conventional TEM, cytochemical and immunolabelling techniques.

Practice: Students will focus their pratical experience in basic transmission electron microscopy techniques using biological samples. Specimens will be fixed, dehydrated and embedded in Epoxy resin. Hardened resin blocks will be cut by students with a ultramicrotome equipped with a diamond knife. Ultrathin sections will be collected on formvar/carbon coated copper grids, double stained with uranyl acetate and lead citrate and finally observed with a Jeol 100 Sx transmission electron microscope. Students will discuss the results of their own samples together with teacher. The discussion of electron micrographs of different biological samples is also planned.

Lesson: Overview on principles and potential applications of Comet assay and Cytokinesis-block micronucleus Cytome assay for the evaluation of DNA primary and chromosomal damage, respectively..