MACS Academy

T cell flow cytometry – Analyzing antigen-specific T cells extra- and intracellularly

T cells play an essential role in the immune system, as they are responsible for initiation and regulation of immune responses to pathogens. T cells are a heterogeneous cell population, comprising a multitude of subpopulations with distinct functions and phenotypes. Flow cytometry is a widely used method for cell analysis and is often used to elucidate the functions and roles of T cells in health and disease. However, the detection of antigen-specific T cells, for example, is hampered by their very low frequency.

This course provides hands-on experience for detecting and analyzing antigen-specific and non-specific human T cell subsets. Participants will be introduced to immunomagnetic pre-enrichment as a simple method enabling the reliable detection and analysis of rare cell types.

The training covers all subjects from a profound introduction via hands-on training through to comprehensive trouble-shooting.

Agenda

Two different technical approaches will be shown and performed in the lab:

  1. Enrichment of antigen-specific CD4+ T cells by using an activation marker in combination with intracellular cytokine staining
  2. Analysis and enrichment of viable cytokine producing antigen-specific T cells using Cytokine Secretion Assay technology

Course lectures and exercises include:

  • Introduction to rare cell and T cell analysis
  • Introduction to immunomagnetic cell enrichment
  • Antigen-specific T cell cytometry

Laboratory practicals include:

  • Immunofluorescent labeling of cells (surface and intracellular)
  • Immunomagnetic labeling and magnetic cell separation
  • In vitro stimulation of leukocytes
  • Flow cytometric detection and characterization of antigen-specific T cell populations

Learning objectives

  • Identify and overcome the issues encountered in rare cell analysis
  • Be able to perform immunomagnetic cell separation
  • Practice flow cytometric data analysis of T cell populations

Participants should be able to independently perform flow cytometric experiments.

MACS Academy location and training dates

Bergisch Gladbach/Cologne, Germany

San Diego, CA, USA

  • New courses will be available in 2019 - Please contact MACS Academy for available places

Training contacts