Numbers matter: Quantitative and dynamic analysis of the formation of an immunological synapse using imaging flow cytometry

Fariyal Ahmed, Sherree Friend, Thaddeus C. George, Natasha Barteneva, Judy Lieberman

Research output: Contribution to journalArticlepeer-review

27 Citations (Scopus)

Abstract

Activation of T lymphocytes by antigen-presenting cells (APC) results in the formation of an immunological synapse. Following contact with the target cell, key signaling and adhesion molecules polarize within minutes to hours to the T cell-APC interface. Multispectral imaging flow cytometry, a new technology which combines flow cytometry with imaging, was used to visualize and quantify the recruitment of the CD3ε and Lck signaling molecules during the evolution of an immune synapse. Using this technology, thousands of T cell/macrophage conjugates could be analyzed for each experimental time point. Following Ca++ triggered T cell activation, the dynamics of Lck and CD3ε recruitment to the synapse, analyzed by two independent methods, were comparable. However, CD3ε exhibited longer residence times (> 8 min) at the synapse than Lck.

Original languageEnglish
Pages (from-to)79-86
Number of pages8
JournalJournal of Immunological Methods
Volume347
Issue number1-2
DOIs
Publication statusPublished - Aug 15 2009

Keywords

  • Flow cytometry
  • Fluorescence microscopy
  • Imaging flow cytometry
  • Immunological synapse

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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