Nuclear translocation kinetics of NF-[kappa]B in macrophages challenged with pathogens in a microfluidic platform

We have developed a microfluidic platform for real-time imaging of host-pathogen interactions and cellular signaling events. Host cells are immobilized in a controlled environment for optical interrogation of the kinetics and stochasticity of immune response to pathogenic challenges. Here, we have q...

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Bibliographic Details
Published in:Biomedical microdevices 2009-06, Vol.11 (3), p.693-700
Main Authors: James, Conrad D, Moorman, Matthew W, Carson, Bryan D, Branda, Catherine S, Lantz, Jeffrey W, Manginell, Ronald P, Martino, Anthony, Singh, Anup K
Format: Article
Language:English
Subjects:
Biomedical research
Immunology
Kinetics
Real time
Signal transduction
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Summary:We have developed a microfluidic platform for real-time imaging of host-pathogen interactions and cellular signaling events. Host cells are immobilized in a controlled environment for optical interrogation of the kinetics and stochasticity of immune response to pathogenic challenges. Here, we have quantitatively measured activation of the toll-like receptor 4 (TLR4) pathway in RAW264.7 murine macrophage-like cells. This was achieved by measuring the cytoplasm-to-nucleus translocation kinetics of a green fluorescent protein fusion construct to the NF-[kappa]B transcription factor subunit RelA (GFP-RelA). Translocation kinetics in response to live bacteria and purified lipopolysaccharide (LPS) challenges were measured, and this work presents the first demonstration of live imaging of host cell infection on a microfluidic platform with quantitative analysis of an early (
ISSN:1387-2176
1572-8781
DOI:10.1007/s10544-008-9281-5