Protocol

Mast Cells

Volume 1220 of the series Methods in Molecular Biology pp 347-363

Date:

Real-Time Imaging of Ca2+ Mobilization and Degranulation in Mast Cells

  • Roy CohenAffiliated withBaker Institute for Animal Health, Cornell University College of Veterinary Medicine
  • , David A. HolowkaAffiliated withDepartment of Chemistry and Chemical Biology, Cornell University
  • , Barbara A. BairdAffiliated withDepartment of Chemistry and Chemical Biology, Cornell University Email author 

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Abstract

Mast cells play a key role in allergy and inflammation processes as part of the immune response. The activation of mast cells via antigen binding and cross-linking of IgE receptors initiates the onset of dramatic calcium (Ca2+) mobilization dynamics that promote the release of mediators of inflammation and allergy. Ca2+ signaling in mast cells has been studied extensively using a variety of research tools and techniques. In these studies, a large number of proteins have been identified to participate in various stages of these processes.

Here we describe single-cell imaging as an important approach for examining Ca2+ signaling and exocytosis in mast cells. Single-cell imaging tools have advanced significantly over the last 10 years, in part due to improvements in microscope technology and in part due to the development of a new generation of Ca2+ indicators and genetically encoded Ca2+ sensors. The single-cell imaging techniques described here provide the spatial and temporal resolution required to decipher the signaling events that are critical for mast cell functions.

Key words

Live-cell imaging Calcium (Ca2+) signaling Calcium dynamics Mast cell degranulation