The course aims to give students knowledge of flow cytometric instrumentation, functioning and application to several aspects of cell biology. It will provide the basic information necessary to better understand diagnostic cytometry. In particular, the methodologies to study the cell survival and death (apoptosis and necrosis), the cell cycle, the immunophenotyping, the cell proliferation and differentiation and some cell functions. After the acquisition of the basic information regarding data analysis, the flow cytometric approach will be compared with the standard methodologies of microscopy and biochemistry.

Notes on the structure and functioning of flow cytometers. The fluidic, the optic and the electronic systems. Light scattering, fluorescence, and fluorochromes. Generation of cytometric graphs and interpretation of flow cytometric data. Notes on innovative application of cytometry, such as flow cytometric cell sorting, mass cytometry and image cytometry. Use of controls in flow cytometry. Analyses of cell survival and death. Evaluation of cell apoptosis and necrosis. Apoptosis/necrosis progression. Analysis of DNA content. Analysis of cell cycle with mono-parametric (propidium iodide, PI) and bi-parametric (PI/BrdU) approaches. Evaluation of the frequency of cell mutants by flow cytometry using the X-linked Pig-a reporter gene involved in paroxysmal nocturnal haemoglobinuria. Analysis of phenotype and function of the immune system. Analysis of intracytoplasmic proteins (cytokines and onco-proteins) and of cytotoxic activity of some cells of peripheral blood samples. Haematopoietic stem cell biology. Absolute count of haematopoietic progenitors. Haematopoietic differentiation in vitro. Use of monoclonal antibodies against surface antigens in the identification of the lympho-haematopoietic subpopulations of peripheral blood and in the description of haematopoietic cell differentiation.

D1-KNOWLEDGE AND COMPREHESION  At the end of the course, students  must prove to have learned and understood program content.

D2 ABILITY TO APPLY KNOWLEDGE AND COMPREHENSION  Students must be able to describe the various methodologies and know when to apply  them in different contexts.

D3 CRITICAL THINKING SKILLS Students must have a sufficient command of what they have learned to correctly and critically establish the correlations between the methodologies studied and the information that can be derived from these; they must be able to use these skills to solve scientific questions proposed.

D4 COMMUNICATION SKILLS   Students must show competence when relating their knowledge and must use appropriate terminology. Students must be able to communicate what they have learned clearly and comprehensibly even to those who have no knowledge of the topic.

D5- LEARNING SKILLS  Students must be able to examine and comprehend scientific texts so as to become actively involved in developing their knowledge of the topic of study. 

The teaching material prepared by the lecturer in addition to recommended textbooks (such as for instance slides, lecture notes, exercises, bibliography) and communications from the lecturer specific to the course can be found inside the Moodle platform › blended.uniurb.it

Lessons in laboratory with data analyses with cytofluorimetric softwares.

Teaching

The course will consists in lecturers and lab experiences.

Course books

Lecture notes of Citometria applicata. L.Zamai et al., available in Moodle; slides of the lessons in pdf format (WEB page) and scientific articles

Assessment

Oral examination (can be either in English or Italian). To verify the level of knowledge, it will be asked at least 3 questions regarding the topics of the course. The evaluation will be performed on the basis of the knowledge, the critical approach, the speaking ability and appropriate terminology to present the topics. Each of the evaluation criteria is judged on 4 level scale. Particular attention will be paid to the first two criteria.

Disability and Specific Learning Disorders (SLD)

Students who have registered their disability certification or SLD certification with the Inclusion and Right to Study Office can request to use conceptual maps (for keywords) during exams.

To this end, it is necessary to send the maps, two weeks before the exam date, to the course instructor, who will verify their compliance with the university guidelines and may request modifications.

Teaching

WEB page with lessons in pdf format and scientific articles

Course books

Lecture notes of Citometria applicata. L.Zamai et al., available in Moodle; slides of the lessons in pdf format (WEB page) and scientific articles

Assessment

Oral examination (can be either in English or Italian). To verify the level of knowledge, it will be asked at least 3 questions regarding the topics of the course. The evaluation will be performed on the basis of the knowledge, the critical approach, the speaking ability and appropriate terminology to present the topics. Each of the evaluation criteria is judged on 4 level scale. Particular attention will be paid to the first two criteria.

Disability and Specific Learning Disorders (SLD)

Students who have registered their disability certification or SLD certification with the Inclusion and Right to Study Office can request to use conceptual maps (for keywords) during exams.

To this end, it is necessary to send the maps, two weeks before the exam date, to the course instructor, who will verify their compliance with the university guidelines and may request modifications.

The student can request to sit the final exam in English.