Resolving dynamics of cell signaling via real-time imaging of the immunological synapse.

Technical Report · Thu Oct 01 00:00:00 EDT 2009

DOI:https://doi.org/10.2172/974405· OSTI ID:974405

Stevens, Mark A; Pfeiffer, Janet R ^[1]; Wilson, Bridget S ^[1]; Timlin, Jerilyn Ann; Thomas, James L ^[1]; Lidke, Keith A ^[1]; Spendier, Kathrin ^[1]; Oliver, Janet M ^[1]; Carroll-Portillo, Amanda ^[1]; Aaron, Jesse S; Mirijanian, Dina T; Carson, Bryan D; Burns, Alan Richard; Rebeil, Roberto

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1. University of New Mexico, Albuquerque, NM

This highly interdisciplinary team has developed dual-color, total internal reflection microscopy (TIRF-M) methods that enable us to optically detect and track in real time protein migration and clustering at membrane interfaces. By coupling TIRF-M with advanced analysis techniques (image correlation spectroscopy, single particle tracking) we have captured subtle changes in membrane organization that characterize immune responses. We have used this approach to elucidate the initial stages of cell activation in the IgE signaling network of mast cells and the Toll-like receptor (TLR-4) response in macrophages stimulated by bacteria. To help interpret these measurements, we have undertaken a computational modeling effort to connect the protein motion and lipid interactions. This work provides a deeper understanding of the initial stages of cellular response to external agents, including dynamics of interaction of key components in the signaling network at the 'immunological synapse,' the contact region of the cell and its adversary.

• Research Organization: Sandia National Laboratories (SNL), Albuquerque, NM, and Livermore, CA (United States)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC04-94AL85000
• OSTI ID: 974405
• Report Number(s): SAND2009-6396; TRN: US201009%%70
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE
BACTERIA
LIPIDS
MACROPHAGES
MAST CELLS
MEMBRANES
MICROSCOPY
PROTEINS
REFLECTION
SIMULATION
SPECTROSCOPY
Microscopy
Electron-instrumentation.
Protein-analysis.
Microscopy-Technique.