OVERVIEW

Flow cytometry is a widely used analytical method in research laboratories and not only.  It allows to analyze a large number of cells in a short time by quantifying multiple parameters for each cell. In the field of biological research, flow cytometry has become fundamental, enabling a major advance in understanding cellular dynamics and cellular communication. 

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

TEACHING METHODS

Lessons and discussion 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

TEACHERS AND EXAM BOARD

Exam Board

DANIELA FENOGLIO (President)

ALESSIA PARODI

LESSONS

Class schedule

The timetable for this course is available here: Portale EasyAcademy