AIMS AND CONTENT

AIMS AND LEARNING OUTCOMES

The Immunohistochemistry course consists of the following macro topics and their respective training objectives.

1. IMMUNISTOCHEMICAL TECHNIQUES: this course aims to provide students with the fundamental notions for the preparation and critical evaluation of immunohistochemical preparations, including internal and external quality controls and understanding the choice of reagents and markers.

At the end of the course the student will be able to:

- Understand the clinical and diagnostic context that leads to the request for an immunohistochemical examination (IHC).
- Understand the principles of immunohistochemical reaction, the different immunological reactions and the detection methodologies (direct, indirect, polymer systems, etc.).
- Identify the different types of antibodies (monoclonal, polyclonal, primary, secondary, etc.), chromogens, detection systems and equipment used.
- Understand the different applications of diagnostic markers.
- Know the different phases of tissue preparation and the influence of each phase on the quality of the reaction result.
- Identify the positive and negative controls needed to validate an immunostaining and correctly interpret the results, recognizing artifacts and false positives/negatives.
- Be aware of the safety regulations relating to the use of equipment and the handling of reagents used in the preparation of samples.

2. ELECTRON MICROSCOPY: this course aims to provide students with a general understanding of the techniques and applications of this technology in the field of Pathological Anatomy.

At the end of the training course, the student will be able to:
- Understand the physical principles underlying both transmission and scanning electron microscopy.
- Recognize the main types of electron microscopes.
- Acquire the methodologies and protocols for sample preparation in relation to the electron microscopy technique used.
- Be aware of the safety regulations relating to the use of equipment and the handling of reagents used in the preparation of samples.

3. BONE PATHOLOGY AND MANAGEMENT OF OSTEOMEDULLARY BIOPSY (BOM):
- understanding the physiology of bone, acquisition of theoretical and practical knowledge useful for arriving at the diagnosis of bone diseases.
- Recognize the structures of the bone by understanding the macroscopic aspect and the treatment of the decalcification process.
- Understand when the Osteomedullar Biopsy is indicated.
- Know how to describe the phases of the procedure and develop the practical skills for the correct management of the BOM.

4. THE CRYOSTAT: METHODOLOGY AND PROBLEMS OF THE INTRAOPERATIVE EXAMINATION:
- Understand the diagnostic role of the intraoperative examination during a surgical operation, acquire the theoretical skills for the analysis of the samples.
- Understand the physical principle of tissue freezing and the reasons why this technique is used for rapid diagnoses as well as the problems that may arise.

5. THE PRE-ANALYTICAL PHASE OF MOLECULAR BIOLOGY:
- Acquire knowledge and practical skills for the correct management of samples before molecular analysis.
- Understand the phases of management of the cytohistological sample that may affect the final result (extraction of nucleic acid DNA and RNA) from acceptance to delivery to the dedicated laboratory.

6. BIO BANKING OF TISSUES:
- Understand the role of the Bio Bank, acquire practical skills by knowing the regulations and guidelines; learn the techniques of collection, processing and conservation of tissues by freezing in liquid nitrogen.
- Management of the data collected by knowing the ethical and legal regulations relating to biological samples.

7. SLIDE/BLOCK STORAGE AND TRACEABILITY IN ANATOMY PATHOLOGY:
- Understand the importance of managing the storage of both slides and paraffin inclusions in the diagnostic process in long-term storage and use for research purposes.
- Know the use of sample traceability systems from arrival in Anatomical Pathology to storage.
- Understand the importance of electronic storage to ensure data integrity and security.

TEACHING METHODS

Frontal lessons, exercises, small group activities, instant polling.

N.B. Students with Specific Learning Disorders (SLD) are invited to contact the teacher and the School/Department disability contact person at the beginning of the course to agree on any teaching methods that take into account individual learning methods (in compliance with the teaching objectives).

SYLLABUS/CONTENT

IMMUNOHISTOCHEMISTRY

I Immunohistochemistry Lesson: The Basics
The following are the topics covered:
- The clinical and diagnostic context that leads to the request for an immunohistochemical test (IHC)
- The reagents used in an IHC reaction
- The different diagnostic markers and their applications
- Use of the different reagents used in an IHC reaction

II Immunohistochemistry Lesson: Detection Techniques
The following are the topics covered:
- The immunohistochemical reaction in peroxidase/phosphatase
- The different reaction techniques
- Reaction techniques used in relation to the markers sought
- Description of the antigenic visualizations

III Immunohistochemistry Lesson: The staining protocol
The following are the topics covered:
- The suitability of the sample - pre-analytical
- The phases of the IHC reaction protocol
- The reaction variables
- The staining protocol
- The phases of the reaction protocol
- The results obtained

IV Lesson of Immunohistochemistry: Automation in Immunohistochemistry
The following topics are covered:
- Main automatic IHC staining tools and how they work.
- Main reagents used and their characteristics
- How to arrange the consumables and samples inside the instruments
- Possible instrumental errors and possible problems during a staining run: what solutions to adopt

V Immunohistochemistry Lesson: How to prepare a reaction protocol
The following are the topics covered:
- The methodology and phases for preparing a new reaction protocol
- Internal and external quality controls
- How to read and apply the information on the Data Sheets

ELECTRONIC MICROSCOPY

Lesson in Electronic Microscopy
The following are the topics covered:
- Physical principles underlying both transmission and scanning electron microscopy
- Main types of electronic microscopes
- Methodologies and protocols for preparing samples in relation to the electronic microscopy technique used
- Safety regulations relating to the use of equipment and the handling of reagents used in preparing samples

BONE PATHOLOGY AND MANAGEMENT OF OSTEOMEDULLARY BIOPSY
The following are the topics covered:
- The physiology of bone
- Macro and microscopy of bone tissue
- The decalcification protocol

THE CRYOSTAT: METHODOLOGY AND PROBLEMS OF THE INTRAOPERATIVE EXAMINATION
The following are the topics covered:
- The diagnostic role of the intraoperative examination and the physical principle on which it is based
- The different phases of the intraoperative examination and description of the instruments used, reagents and stains

THE PRE-ANALYTICAL PHASE OF MOLECULAR BIOLOGY
The following are the topics covered:
- Correct management of the sample before arrival in the laboratory in relation to the molecular biology examination
- Analysis of potential sample treatment errors that can lead to deterioration of cellular DNA and RNA

BIO BANKING OF TISSUES
The following are the topics covered:
- The role of the Bio Bank, regulations and guidelines
- Collection, processing and conservation techniques of tissues preserved in liquid nitrogen
- Management of the data collected (privacy, ethics, legislation)

ARCHIVING SLIDES AND BLOCKS (TRACEABILITY IN PATHOLOGICAL ANATOMY)
The following topics are covered:
- Management of the archive of slides and inclusions in Pathological Anatomy (guidelines and ministerial regulations)
- Traceability Systems in Pathological Anatomy: software and hardware used
- Data protection in traceability systems

RECOMMENDED READING/BIBLIOGRAPHY

Pathological anatomy and cytopathology for TLB edited by A. Fassina and R. Cappellesso, Ed. CLEUP
- Wheater Histology and microscopic anatomy B. Young, J.S. Lowe, A. Stevens, J.W. Heath, Ed. Elsevier Masson
- Pathological Anatomy -The basics- by Luigi Ruco and Aldo Scarpa, Ed. UTET
- Techniques in Pathological Anatomy M. Melis, F. Carpino, U. Di Tondo (no longer in production)
- Italian Caleidoscope: Immunohistochemical Techniques. No longer in production but downloadable at the following link
- Guidelines for Traceability, Collection, Transport, Conservation and Archiving of cells and tissues for diagnostic investigations of PATHOLOGICAL ANATOMY (retrievable at the following link)

TEACHERS AND EXAM BOARD

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

Third year First semester

Class schedule

The timetable for this course is available here: Portale EasyAcademy

EXAMS

EXAM DESCRIPTION

Written + Oral.

Note: During the exam, the rules regarding the evaluation method of students certified as DSA or with other educational needs indicated by the university will be respected. (e.g.: additional time in the case of written exams, use of concept maps to be viewed for approval by the teachers at least one week before the exam date)

ASSESSMENT METHODS

Open questions with written answer and oral presentation.
Please note that the basic elements of the subject must be known in order for the exam to be considered sufficient.