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Flow cytometry

Description

  • To understand the principles of flow cytometry, its potentialities and limits
  • To get a good overview of possible applications of flow cytometry

Durée:

1 jour

Public

Professionnals from the biotechnology sector : project leaders, researchers, technicians, students, teachers, ...

Introduction to flow cytometry : principles and applications

Compétences développées

•    To understand the principles of flow cytometry, its potentialities and limits
•    To get a good overview of possible applications of flow cytometry

Objectifs de la formation
  • To understand the principles of flow cytometry, its potentialities and limits
  • To get a good overview of possible applications of flow cytometry
Contenu & programme

Introduction to flow cytometry
Fluorescence and cell staining
Instrument overview :

  • Components of the flow cytometer : fluidics, optics and electronics systems
  • Measurement of parameters : FSC, SSC, fluorescence


Data analysis :

  • Graphical representation
  • Gating
  • Compensations

Applications of flow cytometry
Possible fields of use (examples taken from the biomedical sector: fundamental research, pharmacology, medicine, industry,…) :

  • Phenotyping (intra- and extracellular staining)
  • Cell death
  • Cell cycle and proliferation
  • Cell signaling


Flow cytometry derived techniques:

  • Cell sorting
  •  « Multiplex »
  • Multispectral imaging by flow cytometry
Méthode pédagogique

90% theory - 10% demonstrations

Pré-requis

Basic knowledge in cell biology

A practical introduction to flow cytometry

Compétences développées

People with few or no practical experience of flow cytometry will get the opportunity to perform/follow a flow cytometry experiment (multicolor cell phenotyping), from the samples preparation to the interpretation of the results

Objectifs de la formation

People with few or no practical experience of flow cytometry will get the opportunity to perform/follow a flow cytometry experiment (multicolor cell phenotyping), from the samples preparation to the interpretation of the results

Contenu & programme

To perform a multicolor cell phenotyping experiment, including:

  • Experimental strategy
  • Samples preparation
  • Samples analysis on the flow cytometer (demonstration)
  • Detectors and compensations settings
  • Graphical representation and gating strategy
  • Interpretation of results
Méthode pédagogique

60% practicals – 40% demonstrations

Pré-requis

Basic knowledge in cell biology, basic knowledge of flow cytometry or prior attendance to the course « Introduction to flow cytometry and its applications»

A focus on cell phenotyping by flow cytometry

Compétences développées

People with a basic practical experience of flow cytometry will acquire the necessary knowledge to design a customized protocol for multicolor cell phenotyping as well as notions of standardization and “good practices” in flow cytometry

Objectifs de la formation

People with a basic practical experience of flow cytometry will acquire the necessary knowledge to design a customized protocol for multicolor cell phenotyping as well as notions of standardization and “good practices” in flow cytometry

Contenu & programme

Design of a customized protocol for multicolor cell phenotyping

  • Cell preparation
  • Staining strategy (choice of antibodies, choice of fluorophores,…)
  • Experimental conditions (antibody titration, choice of controls,…)
  • Detectors and compensation settings
  • Gating and analysis strategy

Standardization and “good practices” in flow cytometry

  • Standardization of the cytometer
  • Standardization of a protoco
  • Quality control and follow-up
Méthode pédagogique

80% theory - 20% exercices

Pré-requis

Basic knowledge in cell biology, basic practical experience in flow cytometry, or prior attendance to the course: « Introduction to flow cytometry and its applications» and « A practical introduction to flow cytometry »