CODE 68484 ACADEMIC YEAR 2024/2025 CREDITS 2 cfu anno 3 TECNICHE DI LABORATORIO BIOMEDICO 9293 (L/SNT3) - GENOVA SCIENTIFIC DISCIPLINARY SECTOR MED/46 LANGUAGE Italian TEACHING LOCATION GENOVA SEMESTER 1° Semester MODULES Questo insegnamento è un modulo di: PATHOLOGIC ANATOMY AND IMMUNOHISTOCHEMICAL (IHC) TECHNIQUES TEACHING MATERIALS AULAWEB OVERVIEW This course is designed at training future TSLB through theoretical and practical training in the laboratory AIMS AND CONTENT LEARNING OUTCOMES Histochemical and immunohistochemical technique on normal and pathological tissues. Cryostat and molecular biology techniques applied to pathological anatomy AIMS AND LEARNING OUTCOMES The general educational objective is the in-depth knowledge of the staging of the most common and highest incidence neoplasms in the population. Consequently, diagnostic pathways are developed which, from cytology to diagnostic excisional biopsy up to surgical exeresis, characterize the typology of material that reaches Pathological Anatomy. On this basis, aspects of ancillary techniques are developed, such as immunohistochemistry in the definition of receptor structures, characterization of oncogenes and tumor suppressor genes and biomolecular techniques. In this context, two aspects are particularly important: intraoperative diagnostics and fine needle aspiration cytology. TEACHING METHODS classroom lessons with interaction with students SYLLABUS/CONTENT YEAR III SEMESTER I Degree Course in Biomedical Laboratory Techniques TOTAL CFU Pathological Anatomy and Immunohistochemical Techniques - Biomolecular Techniques in Pathological Anatomy 6 Scientific-disciplinary sectors Credits Pathological Anatomy II (Valerio Vellone, Luca Mastracci, Roberto Fiocca, Federica Grillo) 3 Immunohistochemical techniques ( Mariangela Lanata, Giorgia Anselmi) 2 Biomolecular techniques in Pathological Anatomy (Valerio Vellone) 1 Coordinator of the Integrated Course: Valerio Vellone General objectives of the Integrated Course: to acquire the notions and methodologies of immunohistochemical techniques on tissues or other biological material. The individual integrated courses have been divided into topics with a criterion of propaedeuticness and increasing technical difficulty. The immunohistochemical technique is described and explained in detail so that it can be adopted by the student who thoroughly understands the mechanisms and reactions. In this way, methodological approaches and proactive attitudes will be developed in the student, thus outlining their own technical characterization and professional responsibility. The final objective is to achieve a high preparation of immunohistochemical technique, of extemporaneous tests (cryostat) to allow to reasonably envisage modifications, variants and technical adaptations to deal with the problems that may arise in a specialized laboratory. In the face of each technical problem, the teacher will have to precede or follow general concepts of histopathology and cytopathology as well as macro and microscopic anatomo-pathological demonstrations aimed at the global and culturally complete preparation of the student Cultural sphere General theme Didactic Unit Elementary Aims Level Knowledge Type of Competence Skill Type of Didactic Activity Pathological Anatomy Preparation of particular tissues (bone, bone marrow biopsy, etc.) Bone decalcification techniques for traditional inclusions. Special fixation techniques. Particular inclusions without bone decalcification. The section of the bone on a microtome or on a resin microtome.MED46 C 1 1 1 L Pathological Anatomy Particular inclusions (resins) and cutting Inclusion of biopsies and delicate tissues in resin: Fixatives: The inclusion technique: Cutting: particular colours.MED46 C 1 1 1 L/T Pathological Anatomy Preparation of preparations for extemporaneous diagnostics (examination in the freezer) freezing techniques. The freezing times. The characteristics of the various tissues to be frozen. The frozen microtome. Artifacts from inadequate freezing. Microtome section techniques. The basic quick colors. MED46 CF 2 2 3 L/T Pathological Anatomy Basic immunohistochemical techniques Definition of poly and monoclonal antibodies. The preparation of the tissues. Microtome sections for immuno. Internal and external controls. Immunohistochemical detection techniques. Unmasking of antigenic sites. Peroxidase inhibition problem. Concept of sensitivity and specificity of the method. Manual, semi-automatic and automatic systems and the choice of chromogens.MED46 CF 2 2 3 L/T Pathological Anatomy Immuno-fluorescence techniques Fixation of the material for immunofluorescence. The preparation of the sections of cryostatted or fixed and included material. The coloring. Principles of the fluorescence microscope. Assembly of sections. In liquid phase. MED46 C 1 1 1 L/T Pathological anatomy Electron microscopy techniques Transmission and scanning electron microscopy. The levy. The fixation. Inclusion. Techniques for microtome sections. Coloring. Staining of semithin sections for light microscopy. Metallization for scanning microscopy. MED46 C 1 1 1 L/T Pathological anatomy Cytogenetic techniques Study of mitosis. Study of chromosomes. Methods of staining of chromosomes. Flow cytometry: from principles to applications. MED46 C 1 1 1 L/T Pathological Anatomy Molecular biology techniques in Preanalytical Pathological Anatomy, in situ hybridization, FISH, PCR and tissue sequencing.MED46 C 1 1 1 L/T Pathological Anatomy Archiving and conservation of material Concept of archiving and cataloging of material. Legislation on material retention times. biobanks. Storage of frozen tissue samples. Cryopreservation techniques. MED46 CF 2 2 2 L/T RECOMMENDED READING/BIBLIOGRAPHY Cardillo MR – Nozioni di tecniche diagnostiche di Anatomia Patologica – Antonio Delfino Editore, Roma, 2009. Ruco L, Scarpa A – Anatomia Patologica, Le basi. – UTET, Torino,2007. TEACHERS AND EXAM BOARD MARIANGELA LANATA Ricevimento: Request information using this email: mariangela.lanata@galliera.it Galliera hospitals S.C. Analysis Laboratory GIORGIA ANSELMI Exam Board ROBERTO FRANCO FIOCCA (President) FEDERICA GRILLO (President) LUCA MASTRACCI (President) GIORGIA ANSELMI MARIANGELA LANATA VALERIO GAETANO VELLONE (President and Coordinator of Integrated Course) LESSONS LESSONS START III year I semester Class schedule IMMUNOHISTOCHEMICAL (IHC) TECHNIQUES EXAMS EXAM DESCRIPTION 30 triplets of mixed questions concerning technical subjects of histology, cytology and subjects of a general nature of histocytopathology were previously prepared. In practice, each sector of the educational program was separated into questions and subsequently the questions were randomly divided into triplets. Each triplet has been ordered with progressive numbering. An alphabetical letter is extracted which, placing the candidates in alphabetical order, marks the beginning of the exams. Each candidate, at the time of the exam, receives a typewritten sheet with a triplet of questions; the first candidate to start the exam is the one whose surname initial corresponds to the letter drawn. The candidate has 15 minutes to read the questions and prepare a list of answers. The exam is carried out separately by at least two teachers, each of whom ask a question. At the end of the exam, the teachers decide the vote that the candidate can accept or refuse. ASSESSMENT METHODS The exam verifies the achievement of the expected learning outcomes: ability to apply knowledge through the description of the solution to a problem, and communication skills, on the basis of the scheme that has been added in the program to each topic covered. For each topic, it will be verified the level of knowledge, the type of competence required and the ability necessary to relate to the specific topic. Ultimately, the candidate has to be able to demonstrate the achievement of the strategic purpose in terms of knowing, knowing how to do, knowing how to be.