PRINCIPLES OF FLOW CYTOMETRY

iten
Code
96332
ACADEMIC YEAR
2021/2022
CREDITS
2 credits during the 2nd year of 11158 APPLIED AND EXPERIMENTAL BIOLOGY (LM-6) GENOVA
SCIENTIFIC DISCIPLINARY SECTOR
MED/09
TEACHING LOCATION
GENOVA (APPLIED AND EXPERIMENTAL BIOLOGY)
semester
2° Semester
Teaching materials

OVERVIEW

Flow cytometry is a very flexible platform for the characterization of cells and particles in suspension, which allows for the simultaneous analysis of physical parameters and fluorescence characteristics to address the complexity of biological systems. Due to its versatility and power, cytometry is widely applied to the study and knowledge in many areas of biology and medicine, therefore very widespread also in the laboratories of pharmaceutical companies.

AIMS AND CONTENT

LEARNING OUTCOMES

The course aims at providing basic training with a broad dissemination technique
In particular, the course aims at achieving the following objectives:
- knowledge of the principles on which flow cytometry and instrumentation are based;
- learning of basic techniques;
- deepening of some applications of common use;
- reading and interpreting the obtained results

AIMS AND LEARNING OUTCOMES


Attendance and participation in training activities (lectures and laboratory attendance) will allow the student to acquire the necessary knowledge to deal with the different types and approaches that apply cytometric measurements in the biological field.
Specifically, the course aims to achieve the following objectives:
- knowledge of the fundamental principles of flow cytometry and instrumentation; - reference points and guidelines in the execution of cytometric measurements; - deepening of applications for studies in the biological field; - reading and interpretation of the results obtained.

PREREQUISITES

To effectively deal with the contents of the course, the following basic knowledge is required: structure of the cell and its components, knowledge of the cell cycle, general notions of light-matter interaction and the phenomenon of fluorescence.

TEACHING METHODS

The course includes 16 hours of lectures delivered through multimedia presentations and presentation of cytometric files.

 

SYLLABUS/CONTENT

Basic principles of fluorescence and light-particle interaction

Basic principles of cytometry: General concepts underlying the operation of the flow cytometer and what are the main optical parameters measurable in cytometry

Instrumentation: - Optical part, fluidic part, electronic part

Instrumentation - part of setting: Instrumentation control procedures (calibration, calibration, linearity); Setting the threshold of analysis, compensation

Sample preparation: basic knowledge on the most important techniques for preparing the samples to be analyzed; Fresh, fixed or frozen samples

Fluorescent Cytometry Probes: General information on colorants for nucleic acids, proteins, and the measurement of key cellular features.

Management and Analysis of Generated Data: Principal concepts related to the analysis of a cytometric file: notions about the different gating modes; Off-line compensation of fluorescents; Isotypic and isoclonic control, fluorescence minus one (FMO); Criteria for the assessment of positivity; Main typologies of cytometric data representation.

Cytometric Platform Applications: surface markers, intracytoplasmic and intranuclear markers; Evaluation of cell proliferation; Apoptosis assessment; Cell signaling; cytotoxicity; Polychromatic cytometry; Manipulation of cultured cell lines and their preparation for cytometric assays (with particular reference to adhesion cultures: detachment, fixation, etc.); Treatment and analysis of solid tissue samples (biopsies) with particular reference to the various disaggregation techniques; Microvescicole study

RECOMMENDED READING/BIBLIOGRAPHY


TOOLS SUPPORTING THE DIDACTICS: All the slides used during the lessons will be available on AulaWeb at the end of the series of lessons in PDF format together with other bibliographic material.

TEACHERS AND EXAM BOARD

Office hours: The interview with the teachers can be requested by mail or by telephone daniela.fenoglio@unige.it - telephone 010 / 3533027-3029 daniele.reverberi@hsanmartino.it - telephone 010 / 5558974-8972 (preferably by mail) The teachers' studies are located: Dr. Daniela Fenoglio, at the Direzione of the Center of Excellence for Biomedical Research (CEBR) - Viale Benedetto XV, 7-16132 Genoa Dott. Daniele Reverberi, Molecular Pathology Unit of Policlinico San Martino, Padiglione 15 (IST-sud) - Viale R. Benzi 10 - 16132 Genova ​​

Exam Board

DANIELA FENOGLIO (President)

ALESSIA PARODI

LESSONS

TEACHING METHODS

The course includes 16 hours of lectures delivered through multimedia presentations and presentation of cytometric files.

 

LESSONS START

The lessons of the first semester will start from September 30th 2019 and will end by January 17th 2020, the lessons of the second semester will start from February 17th 2020 and will end by June 12th 2020.
Consult detailed timetable at the following link: https://easyacademy.unige.it/portalestudenti/

EXAMS

EXAM DESCRIPTION

The exam consists of an oral test and takes place in the presence of the Course teachers. The interview lasts about 25 minutes and focuses on the program carried out during the lectures. Through the discussion of various topics, the Commission verifies the achievement of the educational objectives envisaged by the teaching.
At the end of the interview, the teachers of the Commission formulate a final vote which derives from the overall judgment of the candidate.
The grade of 30/30 with honors will be awarded when the knowledge / skills of the subject are excellent

ASSESSMENT METHODS

The oral exam will mainly focus on the topics covered during the lectures and will aim to verify the achievement of the appropriate level of knowledge and in-depth study of the topics covered. In particular, during the interview, the student must demonstrate knowledge of: the anatomy of a cytometer and the fundamental principles of flow cytometry, the characteristics of the main fluorochromes, the sample preparation protocols for the different types of cytometric measurements, the guidelines in the acquisition and analysis of samples, the main applications of the cytometric platform in biology and medicine.

FURTHER INFORMATION


Regular class attendance is strongly recommended.