control and manipulation of pathogens with an optical trap for live cell imaging of intercellular interactions控制和操纵与活细胞成像的光学陷阱病原体细胞间相互作用.pdf

Control and Manipulation of Pathogens with an Optical Trap for Live Cell Imaging of Intercellular Interactions 1 2 3 1 3 Jenny M. Tam , Carlos E. Castro , Robert J. W. Heath , Michael L. Cardenas , Ramnik J. Xavier , 2 1 Matthew J. Lang , Jatin M. Vyas * 1 Division of Infectious Disease, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, United States of America, 2 Mechanical Engineering and Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America, 3 Center for Computational and Integrative Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, United States of America 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. Hig