Control and Manipulation of Pathogens with an Optical Trap for Live Cell Imaging of Intercellular Interactions
Name
Tam-2010-Control and Manipulation of Pathogens with an Optical Trap for Live Cell Imaging of Intercellular Interactions.pdf
Size
597.17 KB
Format
Adobe PDF
Checksum (MD5)
b6761e738adf19b60567d1415c9b0dfa
Author(s)
Tam, Jenny M.
Castro, Carlos E.
Heath, Robert J. W.
Cardenas, Michael L.
Xavier, Ramnik J.
Lang, Matthew J.
Vyas, Jatin M.
Date Issued
December 2010
Journal
PLoS ONE
Publisher
Public Library of Science
Citation
Tam, Jenny M. et al. "Control and Manipulation of Pathogens with an Optical Trap for Live Cell Imaging of Intercellular Interactions." PLoS ONE 5(12): e15215.
Version
Final published version
Abstract
The application of live cell imaging allows direct visualization of the dynamic interactions between cells of the immune system. Some preliminary observations challenge long-held beliefs about immune responses to microorganisms; however, the lack of spatial and temporal control between the phagocytic cell and microbe has rendered focused observations into
the initial interactions of host response to pathogens difficult. This paper outlines a method that advances live cell imaging by integrating a spinning disk confocal microscope with an optical trap, also known as an optical tweezer, in order to provide exquisite spatial and temporal control of pathogenic organisms and place them in proximity to host cells, as
determined by the operator. Polymeric beads and live, pathogenic organisms (Candida albicans and Aspergillus fumigatus) were optically trapped using non-destructive forces and moved adjacent to living cells, which subsequently phagocytosed the trapped particle. High resolution, transmitted light and fluorescence-based movies established the ability to observe early events of phagocytosis in living cells. To demonstrate the broad applicability of this method to immunological studies, anti-CD3 polymeric beads were also trapped and manipulated to form synapses with T cells in vivo, and time-lapse imaging of synapse formation was also obtained. By providing a method to exert fine control of live pathogens with respect to immune cells, cellular interactions can be captured by fluorescence microscopy with minimal perturbation to cells and can
yield powerful insight into early responses of innate and adaptive immunity.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Mechanical Engineering
Terms of Use
Creative Commons Attribution
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1371/journal.pone.0015215
