LEARNING OUTCOMES OF THE COURSE UNIT
This course will be mainly focused on the morphologic and molecular techniques applied to the study of pathologic cells and tissues, crucial to provide the pathologist with the best-quality cyto-histologic samples, equipped with all the additional informations necessary to reach the most accurate diagnosis.
This course will lead the students to develop knowledge and understanding of the basic principles for the correct handling and processing of histologic and cytologic samples. Moreover, thorough knowledge and understanding of the basis of the morphologic, histochemical, immunohistochemical, ultrastructural and molecular biology techniques ancillary to pathologic diagnosis will be reached. Training periods in the histopathology laboratories will allow the students to practically apply the acquired theoretical knowledge.
The students will also acquire:
- knowledge of the specific measures for safety and prevention, that will make them able to work in the Pathologic Anatomy laboratories in a mindful way, without causing damage to themselves, to collegues and to the environment;
- the ability to analize in a critical way all the phases of the technical work, reaching a judgement-making capacity that will allow them to contribute to identify and solve possible technical troubles in the laboratory procedures;
- communication skills that will be of paramount importance to estabilish cooperation with collegues and with biologist and pathologists, in order to discuss technical results and propose critical evaluation of the procedures falling within their competence. This will help to improve the quality of the technical work carried out in the pathology laboratory, eventually contributing to improve the final product, i.e. the cyto-histopathologic diagnosis;
- learning skills that will allow them to self-sufficiently follow the evolution of the technological and instrumental progress in the pathology field, in order to understand and, in case, contribute to the introduction of new technical advances. This may be especially relevant for the molecular biology techniques that are becoming more and more essential in the field of oncologic diagnosis and therapy.
Knowledge of the basis of histology and having passed all the first year's exams
COURSE CONTENTS SUMMARY
The first lessons of the course will be focused on the procedures that characterize a Histopathology laboratory, including its organization, software management system, instrumentation and specific measures for safety and damage prevention. All the technical steps undertaken by a histologic sample since its acceptance at the institute of Pathologic Anatomy will be fully explored: they include fixation, processing, paraffin embedding and sectioning of histologic slides. Similarly, the pathway followed by cytologic samples will be taught. Also the rationale and the technical procedures for intraoperative histologic examination by means of frozen section, a rapid test that can influence the course of a surgical operation, will be covered.
The second part of the course will be dedicated to the cyto-histologic stains essential to the identification of cell/tissue components, structures and specific chemical substances, in normal and pathologic conditions. Both the classic histomorphologic/histochemical stains (hematoxylin-eosin, PAS, etc) and the more recent immunohistochemical procedures will be fully covered. Such stains represent a valuable aid to the pathologist to reach the most accurate diagnosis.
The last part of the course will be focused on a heterogeneous group of techniques ancillary to the pathologic diagnosis, namely trasmission electron microscopy and molecular biology techniques (Fluorescent In Situ Hybridization, FISH; Polymerase Chain Reaction, PCR). In particular, molecular methods may allow the identification of genetic alterations in tumoral tissues routinely processed for histopathology (formalin-fixed, paraffin-embedded) or in the DNA extracted from them. These techniques provide the pathologist and the oncologist with paramount informations having potential implications for the diagnosis, prognosis and prediction of response to pharmacologic treatment with biologically-targeted drugs.
AA.VV. Anatomia Patologica: le basi. A cura di L. Ruco, A. Scarpa. UTET Scienze Mediche, Torino, 2007
AA.VV. Manuale di Tecnica Cito-Istologica. A cura di S. Daniel, T. Zanin.
Documentazione Scientifica Editrice (DSE), Bologna, 1997
PDF files of the Power Point slides presented during the course
ASSESSMENT METHODS AND CRITERIA
Achievement of learning outcomes will be verified by means of a final oral exam, whose score will be expressed in thirtieth; the minimal threshold to pass the exam will be 18/30, the best mark will be represented by 30/30 “cum laude”. By means of a series of questions touching a wide variety of topics treated during the course, the global level of knowledge and understanding of the theoretical and practical subjects, as well as the ability to logically think and connect related topics, will be verified.
The course will be based on frontal teaching: attendance to at least 70% of the classes will be mandatory. At the end of each lesson, the PDF file of the Power Point slides will be uploaded and published online in the dedicated university webpage, representing the main didactic material for the exam preparation.
Organization, software management system and instrumentation of the histopathology laboratory
Types of histologic samples: needle biopsies, endoscopic and surgical biopsies, surgical specimens
Handling of histologic samples: acceptance, gross examination, surgical margins staining, sampling
Tissue fixation, processing, paraffin embedding, sectioning
Intraoperative frozen sections examination
Security and damage prevention measures in Pathologic Anatomy
Routine (haematoxylin-eosin), histomorphologic and histochemical stains
Immunohistochemistry: principles and applications to histopathologic diagnosis
Diagnostic cytology: exfoliative (PAP test), liquid, needle aspiration cytology
Preparation of cytological specimens: smear, thin layer, cytocentrifugation; fixation, staining
Optical and electron microscope
Transmission electron microscopy: sample fixation, processing, embedding, sectioning, staining: applications to neoplastic and non-neoplastic pathology
Molecular biology techniques applied to Pathologic Anatomy:
- FISH (Fluorescent In Situ Hybridization): general principles, applications to oncologic pathology (HER2 gene amplification, ALK gene translocation, 1p19q deletion);
- PCR (Polymerase Chain Reaction) / fragment analysis / pyrosequencing: general principles, applications to pathology (clonality analysis in lymphoproliferative disorders, microsatellite instability and KRAS-NRAS /BRAF gene mutational analysis in colo-rectal cancer, MGMT-gene methylation analysis in cerebral neoplasms).